Composition and method for inhibiting platelet aggregation

ABSTRACT

This invention is directed to a method of preventing or treating diseases or conditions associated with platelet aggregation. The method is also directed to a method of treating thrombosis. The method comprises administering to a subject a pharmaceutical composition comprising a therapeutic effective amount of P2Y 12  receptor antagonist compound, wherein said amount is effective to bind the P2Y 12  receptors on platelets and inhibit ADP-induced platelet aggregation. The P2Y 12  receptor antagonist compounds useful for this invention include mononucleoside 5′-monophosphates, mononucleoside polyphosphates, and dinucleoside polyphosphates of general Formula I, or salts thereof. The present invention also provides novel compounds of mononucleoside 5′-monophosphates, mononucleoside polyphosphates, and dinucleoside polyphosphates. The present invention further provides pharmaceutical formulations comprising mononucleoside 5′-monophosphates, mononucleoside polyphosphates, or dinucleoside polyphosphates.

[0001] This application is a continuation-in-part of U.S. applicationSer. No. 09/934,970, filed Aug. 21, 2001, which is acontinuation-in-part of U.S. application Ser. No. 09/643,138, filed Aug.21, 2000. The contents of both applications are incorporated herein byreference in their entirety.

TECHNICAL FIELD

[0002] This invention relates to mono- and dinucleoside polyphosphatecompounds and the method of using such compounds in the prevention ortreatment of diseases or conditions associated with plateletaggregation, including thrombosis in humans and other mammals.

BACKGROUND OF THE INVENTION

[0003] Hemostasis is the spontaneous process of stopping bleeding fromdamaged blood vessels. Precapillary vessels contract immediately whencut; within seconds, thrombocytes, or blood platelets, are bound to theexposed matrix of the injured vessel by a process called plateletadhesion. Platelets also stick to each other in a phenomenon known asplatelet aggregation to form a platelet plug to stop bleeding quickly.

[0004] An intravascular thrombus results from a pathological disturbanceof hemostasis. Platelet adhesion and aggregation are critical events inintravascular thrombosis. Activated under conditions of turbulent bloodflow in diseased vessels or by the release of mediators from othercirculating cells and damaged endothelial cells lining the vessel,platelets accumulate at a site of vessel injury and recruit furtherplatelets into the developing thrombus. The thrombus can grow tosufficient size to block off arterial blood vessels. Thrombi can alsoform in areas of stasis or slow blood flow in veins. Venous thrombi caneasily detach portions of themselves called emboli that travel throughthe circulatory system and can result in blockade of other vessels, suchas pulmonary arteries. Thus, arterial thrombi cause serious disease bylocal blockade, whereas venous thrombi do so primarily by distantblockade, or embolization. These conditions include venous thrombosis,thrombophlebitis, arterial embolism, coronary and cerebral arterialthrombosis, unstable angina, myocardial infarction, stroke, cerebralembolism, kidney embolisms and pulmonary embolisms.

[0005] A number of converging pathways lead to platelet aggregation.Whatever the initial stimulus, the final common event is crosslinking ofplatelets by binding fibrinogen to a membrane binding site, glycoproteinIIb/IIIa (GPIIb/IIIa). Compounds that are antagonists for GPIIb/IIIareceptor complex have been shown to inhibit platelet aggregation (U.S.Pat. Nos. 6,037,343 and 6,040,317). Antibodies against GPIIb/IIIa havealso been shown to have high antiplatelet efficacy (The EPICinvestigators, New Engl. J. Med. (1994) 330:956-961). However, thisclass of antiplatelet agents sometimes causes bleeding problems.

[0006] Thrombin can produce platelet aggregation largely independentlyof other pathways but substantial quantities of thrombin are unlikely tobe present without prior activation of platelets by other mechanisms.Thrombin inhibitors such as hirudin are highly effective antithromboticagents. However, functioning as both antiplatelet and anti-coagulantagents, thrombin inhibitors again can produce excessive bleeding. (TheTIMI 9a investigators, The GUSTO Iia investigators, Circulation, 90:1624-1630 (1994); Circulation, 90: 1631-1637 (1994); Neuhaus K. L. etal., Circulation, 90: 1638-1642 (1994))

[0007] Various antiplatelet agents have been studied for many years aspotential targets for inhibiting thrombus formation. Some agents such asaspirin and dipyridamole have come into use as prophylacticantithrombotic agents, and others have been the subjects of clinicalinvestigations. To date, the powerful agents such as disintegrins, andthe thienopyridines ticlopidine and clopidogrel have been shown to havesubstantial side effects, while agents such as aspirin have useful butlimited effectiveness (Hass, et al., N. Engl. J. Med., 321:501-507(1989); Weber, et al., Am. J. Cardiol. 66:1461-1468 (1990); Lekstrom andBell, Medicine 70:161-177 (1991)). In particular, use of thethienopyridines in antiplatelet therapy has been shown to increase theincidence of potentially life threatening thrombotic thrombocytopenicpurpura (Bennett, C. L. et al N. Engl. J. Med, (2000) 342: 1771-1777).Aspirin, which has a beneficial effect on platelet aggregation (Br. Med.J. (1994) 308: 81-106; 159-168), acts by inducing blockade ofprostaglandin synthesis. Aspirin has no effect on ADP-induced plateletaggregation, and thus has limited effectiveness on platelet aggregation.Furthermore, its well documented high incidence of gastric side effectslimits its usefulness in many patients. Clinical efficacy of some newerdrugs, such as ReoPro (7E3), is impressive, but recent trials have foundthat these approaches are associated with an increased risk of majorbleeding, sometimes necessitating blood transfusion (New Engl. J. Med.(1994) 330:956-961). Thus it appears that the ideal “benefit/risk” ratiohas not been achieved.

[0008] Recent studies have suggested that adenosine 5′-diphosphate(ADP), a common agonist, plays a key role in the initiation andprogression of arterial thrombus formation (Bemat, et al., Thromb.Haemostas. (1993) 70:812-826); Maffrand, et al., Thromb. Haemostas.(1988) 59:225-230; Herbert, et al., Arterioscl. Thromb. (1993)13:1171-1179). ADP induces platelet aggregation, shape change,secretion, influx and intracellular mobilization of Ca⁺², and inhibitionof adenylyl cyclase. Binding of ADP to platelet receptors is requiredfor elicitation of the ADP-induced platelet responses. There are atleast three P2 receptors expressed in human platelets: a cation channelreceptor P2X₁, a G protein-coupled receptor P2Y₁, and a Gprotein-coupled receptor P2Y₁₂ (also referred to as P²Y_(ac) andP2_(T)). The P2X₁ receptor is responsible for rapid calcium influx andis activated by ATP and by ADP. However, its direct role in the processof platelet aggregation is unclear. The P2Y₁ receptor is responsible forcalcium mobilization, shape change and the initiation of aggregation.P2Y₁₂ receptor is responsible for inhibition of adenylyl cyclase and isrequired for full aggregation. (Hourani, et al., The Platelet ADPReceptors Meeting, La Thuile, Italy, Mar. 29-31, 2000)

[0009] Ingall et al. (J. Med. Chem. 42: 213-220, (1999)) describe adose-related inhibition of ADP-induced platelet aggregation by analoguesof adenosine triphosphate (ATP), which is a weak, nonselective butcompetitive P2Y₁₂ receptor antagonist. Zamecnik (U.S. Pat. No.5,049,550) discloses a method for inhibiting platelet aggregation in amammal by administering to said mammal a diadenosine tetraphosphatecompound of App(CH₂)ppA or its analogs. Kim et al. (U.S. Pat. No.5,681,823) disclose P¹, P⁴-dithio-P², P³-monochloromethylene 5′,5^(m)diadenosine P¹, P⁴-tetraphosphate as an antithrombotic agent. Thethienopyridines ticlopidine and clopidogrel, which are metabolized toantagonists of the platelet P2Y₁₂ receptor, are shown to inhibitplatelet function in vivo (Quinn and Fitzgerald, Circulation100:1667-1672 (1999); Geiger, et al., Arterioscler. Thromb. Vasc. Biol.19:2007-2011 (1999)).

[0010] There is a need in the area of cardiovascular and cerebrovasculartherapeutics for an agent that can be used in the prevention andtreatment of thrombi, with minimal side effects, such as unwantedprolongation of bleeding, while preventing or treating target thrombi.

SUMMARY OF THE INVENTION

[0011] This invention is directed to a method of preventing or treatingdiseases or conditions associated with platelet aggregation; suchdiseases include venous thrombosis, thrombophlebitis, arterial embolism,coronary and cerebral arterial thrombosis, unstable angina, myocardialinfarction, stroke, cerebral embolism, kidney embolisms and pulmonaryembolisms. The method is also directed to a method of preventing,treating or reducing the incidence of: thrombosis, thrombotic events,embolic events or pathological conditions associated with such events,where the thrombosis, thrombotic event or embolic event occurs during orafter surgery.

[0012] The method comprises administering to a subject a pharmaceuticalcomposition comprising a therapeutic effective amount of P2Y₁₂ receptorantagonist compound, wherein said amount is effective to bind the P2Y₁₂receptors on platelets and inhibit ADP-induced platelet aggregation.

[0013] The P2Y₁₂ receptor antagonist compounds useful for this inventioninclude compounds of general Formula I, or salts thereof:

[0014] wherein:

[0015] X₁, X₂, and X₃ are independently oxygen, methylene,monochloromethylene, dichloromethylene, monofluoromethylene,difluoromethylene, or imido;

[0016] T₁, T₂, W, and V are independently oxygen or sulfur;

[0017] m=0, 1 or 2;

[0018] n=0 or 1;

[0019] p=0,1, or 2;

[0020] where the sum of m+n+p is from 0 to 5; (monophosphate tohexaphosphate)

[0021] M=H or a pharmaceutically-acceptable inorganic or organiccounterion;

[0022] D₁=O or CH₂;

[0023] B′ is a purine or a pyrimidine residue according to generalFormulae IV and V which is linked to the 1′ position of the furanose orcarbocycle via the 9- or 1-position of the base, respectively;

[0024] Y′═H, OH, or OR₁;

[0025] Z′═H, OH, or OR₂;

[0026] with the proviso that when A=M, at least one of Y′ and Z′ isequal to OR₁ or OR₂ respectively;

[0027] A=M, or

[0028] A is a nucleoside residue which is defined as:

[0029] and which is linked to the phosphate chain via the 5′ position ofthe furanose or carbocycle; wherein:

[0030] D₂=O or CH₂;

[0031] Z═H, OH, or OR₃;

[0032] Y=H, OH, or OR₄;

[0033] with the proviso that at least one of Y′, Z′, Y and Z is equal toOR₁, OR_(2 OR) ₃ or OR₄ respectively;

[0034] B is a purine or a pyrimidine residue according to generalFormulae IV and V which is linked to the 1′-position of the furanose orcarbocycle via the 9- or 1′-position of the base, respectively;

[0035] R₁, R₂, R₃, and/or R₄ are residues which are linked directly tothe 2′- and/or 3′-hydroxyls of the respective furanose or carbocycle viaa carbon atom according to Formula II, or linked directly to two (2′-and 3′-)hydroxyls of the respective furanose or carbocycle via a commoncarbon atom according to Formula II, such that from one to fourindependent residues of R₁, R₂, R₃ and R₄ falling within the definitionof Formula III are present or from one to two independent residues madeup of R+R₂ and/or R₃+R₄ are present.

[0036] The present invention also provides novel mononucleo side5′-monophosphate-, mononucleoside polyphosphate-, and dinucleosidepolyphosphate-compounds which are useful in this invention. The presentinvention further provides pharmaceutical formulations comprisingmononucleoside 5′-monophosphates, mononucleoside polyphosphates, ordinucleoside polyphosphates.

BRIEF DESCRIPTION OF THE FIGURES

[0037]FIG. 1 shows the effect of inhibition of ADP-induced aggregationby different compounds.

DETAILED DESCRIPTION OF THE INVENTION

[0038] This invention is provides a method of preventing or treatingdiseases or conditions associated with platelet aggregation. The methodalso provides a method of treating thrombosis. The method comprisesadministering to a subject a pharmaceutical composition comprising atherapeutic effective amount of P2Y₁₂ receptor antagonist compound,wherein said amount is effective to bind the P2Y₁₂ receptors onplatelets and inhibit ADP-induced platelet aggregation. The P2Y₁₂receptor antagonist compounds useful for this invention include compoundof general Formula I and salts thereof:

[0039] wherein:

[0040] X₁, X₂, and X₃ are independently oxygen, methylene,monochloromethylene, dichloromethylene, monofluoromethylene,difluoromethylene, or imido;

[0041] T₁, T₂, W, and V are independently oxygen or sulfur;

[0042] m=0, 1 or 2;

[0043] n=0 or 1;

[0044] p=0, 1, or 2;

[0045] where the sum of m+n+p is from 0 to 5; (from monophosphate tohexaphosphate)

[0046] M=H, or a pharmaceutically-acceptable inorganic or organiccounterion;

[0047] D═O or CH₂

[0048] B′ is a purine or a pyrimidine residue according to generalFormulae IV and V which is linked to the 1′-position of the furanose orcarbocycle via the 9- or 1-position of the base, respectively;

[0049] Y′═H, OH, or OR₁;

[0050] Z′═H, OH, or OR₂; with the proviso that when A=M, at least one ofY′ and Z′ is OR₁ or OR₂;

[0051] A=M, or

[0052] A is a nucleoside residue which is defined as:

[0053] and which is linked to the phosphate chain via the 5′-position ofthe furanose or carbocycle; wherein:

[0054] D₂=O or CH₂;

[0055] Z═H, OH, or OR₃;

[0056] Y=H, OH, or OR₄;

[0057] with the proviso that at least one of Y′, Z′, Y and Z is equal toOR₁, OR₂, OR₄ or OR₃ respectively.

[0058] B is a purine or a pyrimidine residue according to generalFormulae IV and V which is linked to the 1′ position of the furanose orcarbocycle via the 9- or 1-position of the base, respectively;

[0059] R₁, R₂, R₃, and/or R₄ are residues which are linked directly tothe 2′- and/or 3′-hydroxyls of the respective furanose or carbocycle viaa carbon atom according to Formula II, or linked directly to two (2′-and 3′-)hydroxyls of the respective furanose or carbocycle via a commoncarbon atom according to Formula II, such that from one to fourindependent residues of R₁, R₂, R₃ and R₄ falling within the definitionof Formula II are present or from one to two independent residues madeup of R₁+R₂ and/or R₃+R₄ are present;

[0060] wherein:

[0061] O is the corresponding 2′- and/or 3′-oxygen of the respectivefuranose or carbocycle;

[0062] C is a carbon atom;

[0063] R₅, R₆, and R₇ are H, alkyl, cycloalkyl, aralkyl, aryl,substituted aralkyl, or substituted aryl, such that the moiety definedaccording to Formula II is an ether; or

[0064] R₅ and R₆ are H, an alkyl, cycloalkyl, aralkyl, aryl, substitutedaralkyl, or substituted aryl, and R₇ is alkoxy, cycloalkoxy, aralkyloxy,aryloxy, substituted aralkyloxy, or substituted aryloxy such that themoiety defined according to Formula II is an acyclic acetal or ketal; or

[0065] R₅ and R₆ are taken together as oxygen or sulfur doubly bonded toC, and R₇ is alkyl, cycloalkyl, aralkyl, aryl, substituted aralkyl, orsubstituted aryl, such that the moiety defined according to Formula IIis an ester or thioester; or

[0066] R₅ and R₆ are taken together as oxygen or sulfur doubly bonded toC, and R₇ is amino or mono- or disubstituted amino, where thesubstituents are alkyl, cycloalkyl, aralkyl, aryl, substituted aralkyl,or substituted aryl, such that the moiety according to Formula II is acarbamate or thiocarbamate; or

[0067] R₅ and R₆ are taken together as oxygen or sulfur doubly bonded toC, and R₇ is alkoxy, cycloalkoxy, aralkyloxy, aryloxy, substitutedaralkyloxy, or substituted aryloxy, such that the moiety according toFormula II is a carbonate or thiocarbonate; or

[0068] R₇ is not present and R₅ and R₆ are taken together as oxygen orsulfur doubly bonded to C and both the 2′- and 3′-oxygens of therespective furanose or carbocycle are directly bound to C to form acyclical carbonate or thiocarbonate;

[0069] wherein:

[0070] the O atoms are the 2′- and 3′-oxygens of a furanose orcarbocycle; and the 2′- and 3′-oxygens of the furanose or carbocycle arelinked by a common carbon atom (C) to form a cyclical acetal, cyclicalketal, or cyclical orthoester;

[0071] for cyclical acetals and ketals, R₈ and R₉ are independentlyhydrogen, alkyl, cycloalkyl, aralkyl, aryl, substituted aralkyl,substituted aryl, or can be joined together to form a homocyclic orheterocyclic ring composed of 3 to 8 atoms, preferably 3 to 6 atoms;

[0072] for cyclical orthoesters, R₈ is hydrogen, alkyl, cycloalkyl,aralkyl, aryl, substituted aralkyl, or substituted aryl, R₉ is alkyloxy,cycloalkyloxy, aralkyloxy, aryloxy, substituted aralkyloxy, orsubstituted aryloxy.

[0073] when present, the alkyl, cycloalkyl, aralkyl, aryl, substitutedaralkyl and substituted aryl components of R₅ to R₉ can be generallydefined as, but are not limited to, the following:

[0074] alkyl groups are from 1 to 12 carbons inclusively, eitherstraight chained or branched, with or without unsaturation and with orwithout heteroatoms, are more preferably from 2 to 8 carbonsinclusively, and most preferably 2 to 6 carbons inclusively;

[0075] cycloalkyl groups from 3 to 12 carbons inclusively, morepreferably from 3 to 10 carbons inclusively, and most preferably 3 to 6carbons inclusively, with or without unsaturation, and with or withoutheteroatoms;

[0076] aralkyl groups are from 1 to 8 carbons inclusively in the alkylportion, are more preferably from 1 to 6 carbons inclusively in thealkyl portion, and most preferably are 1 to 4 carbons inclusively in thealkyl portion; as included in the alkyl definition above, the alkylportion of an aralkyl group can include one or more positions ofunsaturation such as a double bonds or a triple bond in the chain whenthe chain includes two or more carbon atoms; the alkyl portion of anaralkyl group can also include one or more heteroatoms and/orsubstituents; the aryl portion of an aralkyl group can be a monocyclicor polycyclic moiety from 3 to 8 carbons inclusively per ring in thearyl portion, more preferably from 4 to 6 carbons inclusively per ring,and most preferably 5 to 6 carbons inclusively per ring; the arylportion of an aralkyl group can can also bear one or more substituentsand/or heteroatoms;

[0077] aryl groups are either monocyclic or polycyclic, are from 3 to 8carbons inclusively per ring, are more preferably from 4 to 6 carbonsinclusively per ring, and are most preferably 5 to 6 carbons inclusivelyper ring; aryl groups can also bear substituents and/or heteroatoms.

[0078] Preferred substituents on the foregoing groups can be, but arenot limited to, hydroxy, nitro, methoxy, fluoro, chloro, bromo, iodo,methyl, ethyl, propyl, butyl, thioalkyl, alkoxy, carboxyl, cyano, amino,substituted amino, trifluoromethyl, phenyl, cyclopropyl, cyclopentyl,and cyclohexyl; and preferred heteroatoms are oxygen, nitrogen, andsulfur.

[0079] When R₅, R₆ and R₇ are not the same, or when R₈ and R₉ are notthe same, a compound according to Formula I can exist in severaldiastereomeric forms. The general structure of Formula I includes alldiastereomeric forms of such materials, when not specified otherwise.Formula I also includes mixtures of compounds of Formula I, includingmixtures of enantiomers, diastereomers and/or other isomers in anyproportion.

[0080] One embodiment of the invention is that A=M, wherein M=H or apharmaceutically-acceptable inorganic or organic counterion. In such anembodiment, the compound can be a nucleoside monophosphate, nucleosidediphosphate, nucleoside triphosphate, nucleoside tetraphosphate,nucleoside pentaphosphate, or nucleoside hexaphosphate with one or bothof the 2′- and/or 3′-positions of the furanose or carbocycle modified.Most preferred are nucleotide monophosphates, nucleotide diphosphates,nucleotide triphosphates, and nucleotide tetraphosphates. When T₂, W, V,or T₁ are sulfur, the preferred position for this atom is on theterminal phosphorous of the polyphosphate chain (i.e. the phosphorousfurthest removed from the nucleoside residue).

[0081] For monophosphates, where m, n, and p are all equal to zero,preferably R₈ is hydrogen and R₉ is aryl or aralkyl, with 1, 2, 3, or 4carbons inclusively in the alkyl portion of an aralkyl group, and 6carbons inclusively in the aryl portion of an aralkyl or aryl group;when the number of carbons in the alkyl portion of an aralkyl group is2, the carbon atoms are most preferably connected by either a double ortriple bond.

[0082] Another embodiment of the invention is that A is a nucleosideresidue defined as:

[0083] and linked to the phosphate chain via the 5′-position of thefuranose or carbocycle (to give a dinucleoside polyphosphate with atleast one of the 2′-, 3′-, 2″- and 3″-positions of the furanose orcarbocycle moieties modified to be OR₁, OR₂, OR₄ or OR₃ respectively).

[0084] When T₂, W, V, and/or T₁ are sulfur, the preferred positions (forsulfur) are T₁ and T₂.

[0085] Further provisions are that when either D₁ or D₂ are oxygen, thecorresponding furanose is preferably in the β-configuration; and thatthe corresponding furanose is most preferably in the β-D-configuration.

[0086] Preferred compounds of general Formula I are molecules whosestructures fall within the definitions of Formula Ia and Formula Ib:

[0087] wherein:

[0088] D₁=O or CH₂;

[0089] D₂=O or CH₂;

[0090] B and B′ are independently purine or pyrimidine residuesaccording to general Formula IV or V;

[0091] m and p=0, 1 or 2; n=0 or 1; such that the sum of m+n+p is from 0to 5, preferably 0 to 4, and most preferably 0 to 3;

[0092] X₁, X₂, and X₃=are independently O, NH, CH₂, CHF, CHCl, CF₂,CCl₂;

[0093] T₁, T₂, V, and W are independently O or S;

[0094] M=H⁺, NH₄ ⁺, Na⁺ or other pharmaceutically-acceptable inorganicor organic counter ion;

[0095] Y′═H, OH, or OR₁;

[0096] Z′═OH or OR₂;

[0097] Z═OH or OR₃;

[0098] Y=H, OH, or OR₄, where R₁, R₂, R₃ and R₄ fall under thedefinition of general Formulae II or III, provided that at least one ofY′, Z′, Z and Y is OR₁, OR₂, OR₃, or OR₄.

[0099] Preferred compounds of Formula Ia include:

[0100] D₁=O or CH₂;

[0101] D₂=0 or CH₂;

[0102] X₁, X₂, and X₃═O;

[0103] T₁, T₂, V, and W═O; or

[0104] D₁=O or CH₂;

[0105] D₂=0 or CH₂;

[0106] X₁ and X₃═O;

[0107] X₂=methylene, monochloromethylene, dichloromethylene,monofluoromethylene, difluoromethylene, or imido;

[0108] T, T₁, T₂, V, and W═O; or

[0109] D₁=O or CH₂;

[0110] D₂=O or CH₂;

[0111] m, n, and p=1; or

[0112] X₁ and X₃═O;

[0113] X₂=methylene, monochloromethylene, dichloromethylene,monofluoromethylene, difluoromethylene, or imido;

[0114] T₁ and T₂=S;

[0115] V and W═O.

[0116] D₁=O or CH₂;

[0117] n and p=0, 1, or 2 such that the sum of n+p is from 0 to 3;

[0118] A=M; wherein M=H⁺, NH₄ ⁺, Na⁺ or otherpharmaceutically-acceptable inorganic or organic counterion;

[0119] B′ is a purine or pyrimidine residue according to generalFormulae IV and V;

[0120] X₁ and X₂ are independently O, NH, CH₂, CHF, CHCl, CF₂, CCl₂;

[0121] T₁, V, and W are independently O or S;

[0122] Y′═H, OH, or OR,

[0123] Z′═H, OH or OR₂, where R₁ and R₂ fall under the definitions ofgeneral Formulae II or III; with the proviso that at least one of Y′ andZ′ is OR, or OR₂, respectively.

[0124] Preferred compounds of Formula Ib include:

[0125] D₁=O or CH₂;

[0126] n and p=0, 1, or 2 such that the sum of n+p is from 0 to 3,preferably 1 to 2;

[0127] X₁ and X₂═O;

[0128] T₁, V, and W═O; or

[0129] D₁=O or CH₂;

[0130] X₁ and X₂═O;

[0131] T₁ and V═O;

[0132] W═S; or

[0133] D₁=O;

[0134] n and p=0 such that the sum of n+p is 0;

[0135] V═O;

[0136] B′ is a purine residue of general Formula IV;

[0137] Y′ and Z′ fall under the definition of general Formula III; or

[0138] D₁=O or CH₂;

[0139] p=0, 1, or 2, n=1, such that the sum of n+p is from 1 to 3;

[0140] X₁═O;

[0141] X₂=methylene, monochloromethylene, dichloromethylene,monofluoromethylene, difluoromethylene, or imido;

[0142] T₁, V, and W═O;

[0143] Y′═H, OH, or OR₁;

[0144] Z′═H, OH or OR₂, where R₁ and R₂ fall under the definition ofgeneral Formulae II or III; with the proviso that at least one of Y′ andZ′ is OR₁ or OR₂, respectively.

[0145] Several preferred compounds also are described by Formula Ib′:

[0146] For compounds of Formula I, B and B′ can independently be apurine residue, as in Formula IV, linked through the 9-position, or apyrimidine residue, as in Formula V, linked through the 1-position. Theribosyl moieties in Formulae Ia, Ib, and Ib′ are in the D-configurationas shown, but can also be L-, or D- and L-. The D-configuration ispreferred for ribosyl moieties.

[0147] wherein:

[0148] R₁₀ and R₁₄ independently are hydroxy, oxo, amino, mercapto,alkylthio, alkyloxy, aryloxy, alkylamino, cycloalkylamino, aralkylamino,arylamino, diaralkylamino, diarylamino, or dialkylamino, where the alkylgroups are optionally linked to form a heterocycle; or

[0149] R₁₀ and R₁₄ independently are acylamino, provided that theyincorporate an amino residue from the C-6 position of the purine or theC-4 position of the pyrimidine; or

[0150] when R₁₀ in a purine or R₁₄ in a pyrimidine has as its first atomnitrogen, R₁₀ and R₁₁ or R₁₄ and R₁₅ can be taken together to form a5-membered fused imidazole ring (to give an etheno compound), optionallysubstituted on the etheno ring with one or more alkyl, cycloalkyl,aralkyl, or aryl moieties, as described for R₅-R₉ above;

[0151] J is carbon or nitrogen, with the provision that when J=nitrogen,R₁₂ is not present;

[0152] R₁₁ is hydrogen, 0 (adenine 1-oxide derivatives) or is absent(adenine derivatives);

[0153] R₁₅ is hydrogen, or acyl (e.g. acetyl, benzoyl, phenylacyl, withor without substituents);

[0154] R₁₂ is hydrogen, alkyl, bromo, azido, alkylamino, arylamino oraralkylamino, alkoxy, aryloxy or aralkyloxy, alkylthio, arythio oraralkylthio, or co-A(C₁₋₆alkyl)B-, wherein A and B are independentlyamino, mercapto, hydroxy or carboxyl;

[0155] R₁₃ is hydrogen, chlorine, amino, monosubstituted amino,disubstituted amino, alkylthio, arylthio, or aralkylthio, where thesubstituent on sulfur contains up to a maximum of 20 carbon atoms, withor without unsaturation, and with or without substituents on the chain;

[0156] R₁₆ is hydrogen, methyl, alkyl, halogen, alkyl, alkenyl,substituted alkenyl, alkynyl, or substituted alkynyl.

[0157] Compounds according to Formulae IV and V where R₁₀ or R₁₄ isacylamino fall within the scope of Formula VI:

[0158] wherein:

[0159] NH is the amino residue at the C-6 position in a purine or theamino residue at the C-4 position in a pyrimidine;

[0160] C is a carbon atom;

[0161] W₁ is oxygen or sulfur;

[0162] R₁₇ is amino or mono- or disubstituted amino, with the aminosubstituent(s) being alkyl, cycloalkyl, aralkyl, or aryl, with orwithout further substituents, unsaturation, or heteroatoms, such thatthe moiety according to Formula VI is a urea or thiourea; or

[0163] R₁₇ is alkoxy, aralkyloxy, aryloxy, substituted aralkyloxy, orsubstituted aryloxy, such that the moiety according to Formula VI is acarbamate or thiocarbamate; or

[0164] R₁₇ is alkyl, cycloalkyl, aralkyl, or aryl, with or withoutsubstituents or heteroatoms, such that the moiety according to FormulaVI is an amide; with definitions of alkyl, cycloalkyl, aralkyl, or arylgroups as previously defined for comparable groups in R₅ to R₉

[0165] The compounds of the present invention can be convenientlysynthesized by those skilled in the art using well-known chemicalprocedures. Mononucloside mono-, di- and triphosphates can be obtainedfrom commercial sources or can be synthesized from the nucleoside usinga variety of phosphorylation reactions which can be found in thechemical literature. Symmetrical and unsymmetrical dinucleotidepolyphosphates can be prepared by activation of a nucleoside mono-, di-or triphosphate with a coupling agent such as, but not limited to,dicyclohexylcarbodiimide or 1,1′-carbonyldiimidazole, followed bycondensation with another nucleoside mono-, di-, or triphosphate, whichcan be the same or different as the activated moiety. Activation ofnucleoside triphosphates with dicyclohexylcarbodiimide gives a cyclicaltrimetaphosphate as the activated species, which can be advantageouslyreacted with a variety of nucleophiles to install unique substituents onthe terminal phosphate of a triphosphate.

[0166] The compounds of the present invention can be prepared byderivatization or substitution at the level of the nucleoside, followedby phosphorylation and condensation as previously described, or thereactions can be carried out directly on the preformed mono- ordinucleotides. In the general Formulae Ia and Ib, the substituents atY′, Z′, Y, and Z can be esters, carbamates, or carbonates, which aregenerally described by Formula II. Esters can be readily prepared byreacting a hydroxyl group of the furanose in a nucleoside or nucleotidewith an activated form of an appropriate organic acid, such as an acidhalide or acid anyhydride in the presence of an organic or inorganicbase. Alternately, use of a suitable coupling reagent such asdicyclohexylcarbodiimide, 1,1′-carbonyldiimidazole and the like toactivate the organic acid can be used to achieve the same result.

[0167] Carbamates or thiocarbamates can be most conveniently prepared byreaction of a hydroxyl group of the furanose in a nucleoside ornucleotide with any of a number of commercially available isocyanates orisothiocyanates, respectively, in an inert solvent. Alternately, when adesired isocyanate or isothiocyanate cannot be obtained from commercialsources, it can be prepared from the corresponding amine by the use ofphosgene or thiophosgene, respectively, or their chemical equivalents.Carbonates or thiocarbonates can be synthesized by reacting the hydroxylgroups of a furanose in a nucleoside or nucleotide with an appropriatehaloformate in the presence of an organic or inorganic base.

[0168] In the general Formulae Ia, Ib and Ib′, the substituents at Y′andZ′, and Y and Z, when taken together, can be taken to mean acetals,ketals or orthoesters, as described in Formula III. Acetals and ketalscan be readily prepared by reaction of the neighboring 2′- and3′-hydroxyl groups of the furanose in an appropriate nucleoside ornucleotide with an aldehyde or ketone, respectively, or their chemicalequivalents, in the presence of an acid catalyst. Particularlyadvantageous is the use of an organic acid, which can effect thetransformation without affecting the integrity of the rest of themolecule. Alternately, strong acids such as trichloroacetic,p-toluenesulfonic, methanesulfonic and the like can be employed incatalytic amounts, in conjunction with inert solvents. Most preferred isformic acid, which can be removed by evaporation under reduced pressure,and is ideally suited to serve as both solvent and catalyst for thesereactions. Alternately, trifluoroacetic acid can be substituted forformic acid in the reaction, provided that the reaction is carried outat low temperatures and the aldehyde or aldehyde equivalent used toprepare the acetal is stable to strong acid conditions. The discovery ofthe utility of formic acid and trifluoroacetic acid for this purpose isone particular aspect of this invention.

[0169] Cyclical orthoesters can be prepared by reaction of theneighboring 2′- and 3′-hydroxyl groups of a furanose with an acylicorthoester, in the presence of an acid. When the nucleoside ornucleotide to be derivatized is a purine that contains a 6-aminofunctionality or is a pyrimidine that contains a 4-amino functionality,it can be converted to the respective urea or thiourea by treatment withan isocyanate or isothiocyanate, respectively, as was previouslydescribed for carbamates or thiocarbamates of the 2′- or 3′-hydroxyls ofa furanose. It was found that reactions of such an amino group withisocyanates or isothiocyanates can be carried out in the presence of oneor more hydroxyl groups on a furanose, by appropriate manipulation ofthe stoichiometry of the reaction.

[0170] All of the derivitization reactions described herein can becarried out on preformed dinucleotide polyphosphates, which results inmultiple products dependent on reaction stoichiometry and on whethermultiple reactive groups are present. When multiple products areobtained, these can be conveniently separated by the use of preparativereverse phase high performance liquid chromatography (HPLC).Particularly advantageous is the use of C 18 or phenyl reverse phasecolumns, in conjunction with gradients that start with ammonium acetatebuffer and end with methanol. The use of a buffer provides fornucleotide stability and improved peak shape of the eluting products andthe use of methanol allows for effective desorption of these lipophiliccompounds from the column. Particularly advantageous is the use ofammonium acetate buffer solutions in conjunction with methanol, as thesesolvents are miscible in all proportions and can be readily removed fromthe chromatographed products by evaporation, followed by lyophilization.

[0171] While separation of multiple products can be done by HPLC,another strategy is to use nucleosides or nucleotides which contain onlya single reactive functionality, whether because only one is present, orby the use of protecting groups to block side reactions at otherpositions in the molecule. This can be done at the level of preformeddinucleotide polyphosphates, or alternately, can be carried out onnucleoside mono-, di-, or triphosphates, leading to novel products intheir own right, or can be coupled to other nucleoside mono-, di, ortriphosphates by the methods which have already been described.

[0172] It will be recognized by those skilled in the art that the abovereactions and purification techniques can also be applied tocarba-ribose analogues (e.g., D₁=CH₂) of nucleosides, nucleotides andtheir derivatives, and that the terms such as “mononucleotide” and“dinucleotide” also apply to the carba-ribose analogues and otherderivatives defined by Formulae I-IV.

[0173] The inventors of the present invention have discovered compoundsthat are antagonists of the effect of ADP on its platelet membranereceptor, the P2Y₁₂ receptor. The compounds provide efficacy asantithrombotic agents by their ability to block ADP from acting at itsplatelet receptor site and thus prevent platelet aggregation. Thus,these compounds can provide a more efficacious antithrombotic effectthan aspirin, but with less profound effects on bleeding thanantagonists of the fibrinogen receptor. Since ADP-induced plateletaggregation is mediated by the simultaneous activation of both P2Y₁₂ andP2Y₁ receptors, the combined administration of the compounds describedhere with antagonists of platelet P2Y₁ receptors could potentiallyprovide a more efficacious antithrombotic effect at concentrations ofeach antagonist that are below the effective concentrations to blockeach receptor subtype in other systems, resulting in a decrease of thepotential manifestation of adverse effects. In addition, these compoundscan be used in conjunction with lower doses of other plateletaggregation inhibitors which work by different mechanisms, to reduce thepossible side effects of said agents. Finally, if the compounds of thepresent invention have sufficient binding affinity and bear afluorescent moiety, they can find uses as biochemical probes for theP2Y₁₂ receptor.

[0174] The compounds of the present invention fall under the definitionof general Formula I which is further divided into general Formulae Ia(dinucleotides), Ib (mononucleotides) and Ib′ (mononucleotidemonophosphates). While potent and selective P2Y₁₂ antagonists can befound within either of these subdivisions, mononucleotides have anadvantage over dinucleotides in terms of ease of synthesis and cost. Ingeneral, diphosphates and triphosphates falling under general Formula Ibare more potent antagonists at P2Y₁₂ than the correspondingmonophosphates of Formula Ib′. However, nucleoside 5′-monophosphates andtheir analogues are easier to prepare and have greater chemical andbiological stability. Thus, for synthetic reasons, a nucleoside5′-monophosphate with appropriate druglike properties is sometimes moreadvantageous than other mononucleotides bearing more than one phosphate,or related dinucleotides.

[0175] Two preferred modifications falling under the definition ofgeneral Formula Ib′ can be made to nucleoside 5′-monophosphates torender them antagonists of the platelet P2Y₁₂ receptor. In general, thepreferred nucleoside 5′-monophosphate starting material chosen for thispurpose is AMP, or an adenosine 5′-monophosphate derivative, as itcontains the appropriate functional groups for the desired modificationsand gives rise to more potent and selective antagonists compared tosimilar modifications of other commonly available nucleotidemonophosphates. The first preferred modification is to install an arylor aralkyl acetal bridging the 2′- and 3′-hydroxyls of the ribose, withthe nature of the aryl or aralkyl group as previously described. Thesecond modification is to add an aminocarbonyl or substitutedaminocarbonyl group to the 6-amino position of the adenine base,resulting in a urea moiety at that position. Substituents on the ureamoiety fall under the definition of R₁₇, as previously described. Theseurea substituents can be broadly categorized as either aromatic oraliphatic in nature. Generally, the most preferred substituent chosenfrom aryl groups is phenyl. When the urea group is an aliphatic urea,the preferred substituents on nitrogen are linear, branched, or cyclic,having from 1 to 6 carbons on the alkyl substituent, and with or withoutunsaturation. More preferred are linear alkyl ureas from 2 to 6 carbonsinclusively, or cyclic alkyl ureas having 3 to 6 carbons in the ring;most preferred are linear alkyl ureas containing from 2 to 4 carbonsinclusively in the chain or cycloalkyl ureas having from 3 to 5 carbonsinclusively in the ring.

[0176] An important aspect of the present invention is that, while anyof the described modifications alone can result in a compound capable ofantagonism of ADP-induced platelet aggregation, it is the combination ofboth 2′/3′ and 6-N modifications in the same molecule that renders thenucleotide a highly potent and selective P2Y₁₂ antagonist.

[0177] Another important aspect of the present invention is the effectof compound structure on the resultant potency in washed plateletsversus potency in whole blood. In general, the potency of a givencompound is lower in whole blood versus washed platelets, ostensibly theresult of increased binding of the compound to the higher levels ofblood proteins in the former. This property is particularly acute fornucleoside 5′-monophosphates versus the corresponding di- andtriphosphates, since there are fewer ionizable groups available in theformer to offset the lipophilic acetal and urea groups which,presumably, increase protein binding in whole blood. Unexpectedly, wefound that compounds containing phenyl ureas exhibited a greater loss ofpotency in whole blood compared to their activity in washed platelets,while those derivatives having aliphatic ureas gave more comparableresults when tested in whole blood and washed platelet assays.Furthermore, compounds with aliphatic ureas were significantly morepotent than their aromatic counterparts in washed platelets. Takentogether, these findings enabled the discovery of the most potent andselective compounds of the present invention.

[0178] The compounds of general Formula I are useful in therapy, inparticular in the prevention of platelet aggregation. The compounds ofthe present invention are thus useful as anti-thrombotic agents, and arethus useful in the treatment or prevention of unstable angina, coronaryangioplasty (PTCA) and myocardial infarction.

[0179] The compounds of the present invention are also useful in thetreatment or prevention of primary arterial thrombotic complications ofatherosclerosis such as thrombotic stroke, peripheral vascular disease,myocardial infarction without thrombolysis.

[0180] Still further indications where the compounds of the inventionare useful are for the treatment or prevention of arterial thromboticcomplications due to interventions in atherosclerotic disease such asangioplasty, endarterectomy, stent placement, coronary and othervascular graft surgery.

[0181] Still further indications where the compounds of the inventionare useful are for the treatment or prevention of thromboticcomplications of surgical or mechanical damage such as tissue salvagefollowing surgical or accidental trauma, reconstructive surgeryincluding skin flaps, and “reductive” surgery such as breast reduction.

[0182] The compounds of the present invention are also useful for theprevention of mechanically-induced platelet activation in vivo such ascardiopulmonary bypass (prevention of microthromboembolism), preventionof mechanically-induced platelet activation in vitro such as the use ofthe compounds in the preservation of blood products, e.g. plateletconcentrates, prevention of shunt occlusion such as renal dialysis andplasmapheresis, thrombosis secondary to vascular damage/inflammationsuch as vasculitis, arteritis, glomerulonephritis and organ graftrejection.

[0183] Still further indications where the compounds of the presentinvention are useful are indications with a diffuse thrombotic/plateletconsumption component such as disseminated intravascular coagulation,thrombotic thrombocytopenic purpura, hemolytic uremic syndrome,heparin-induced thrombocytopenia and pre-eclampsia/eclampsia.

[0184] Still further indications where the compounds of the inventionare useful are for the treatment or prevention of venous thrombosis suchas deep vein thrombosis, veno-occlusive disease, hematologicalconditions such as thrombocythemia and polycythemia, and migraine.

[0185] In a particularly preferred embodiment of the present invention,the compounds are used in the treatment of unstable angina, coronaryangioplasty and myocardial infarction.

[0186] In another particularly preferred embodiment of the presentinvention, the compounds are useful as adjunctive therapy in theprevention of coronary arterial thrombosis during the management ofunstable angina, coronary angioplasty and acute myocardial infarction,i.e. perithrombolysis. Agents commonly used for adjunctive therapy inthe treatment of thrombotic disorders can be used, for example heparinand/or aspirin, just to mention a few.

[0187] A method of treating a mammal to alleviate the pathologicaleffects of atherosclerosis and arteriosclerosis, acute MI, chronicstable angina, unstable angina, transient ischemic attacks and strokes,peripheral vascular disease, arterial thrombosis, preeclampsia,embolism, restenosis or abrupt closure following angioplasty, carotidendarterectomy, and anastomosis of vascular grafts.

[0188] The compounds of this invention can be used in vitro to inhibitthe aggregation of platelets in blood and blood products, e.g. forstorage, or for ex vivo manipulations such as in diagnostic or researchuse. This invention also provides a method of inhibiting plateletaggregation and clot formation in a mammal, especially a human, whichcomprises the internal administration of a compound of Formula (I) and apharmaceutically acceptable carrier.

[0189] Chronic or acute states of hyper-aggregability, such asdisseminated intravascular coagulation (DIC), septicemia, surgical orinfectious shock, post-operative and post-partum trauma, cardiopulmonarybypass surgery, incompatible blood transfusion, abruptio placenta,thrombotic thrombocytopenic purpura (TTP), snake venom and immunediseases, are likely to be responsive to such treatment.

[0190] This invention further provides a method for inhibiting thereocclusion of an artery or vein following fibrinolytic therapy, whichcomprises internal administration of a compound of Formula (I) and afibrinolytic agent. When used in the context of this invention, the termfibrinolytic agent is intended to mean any compound, whether a naturalor synthetic product, which directly or indirectly causes the lysis of afibrin clot. Plasminogen activators are a well known group offibrinolytic agents. Useful plasminogen activators include, for example,anistreplase, urokinase (UK), pro-urokinase (pUK), streptokinase (SK),tissue plasminogen activator (tPA) and mutants, or variants thereof,which retain plasminogen activator activity, such as variants which havebeen chemically modified or in which one or more amino acids have beenadded, deleted or substituted or in which one or more functional domainshave been added, deleted or altered such as by combining the active siteof one plasminogen activator or fibrin binding domain of anotherplasminogen activator or fibrin binding molecule.

[0191] Extracorporeal circulation is routinely used for cardiovascularsurgery in order to oxygenate blood. Platelets adhere to surfaces of theextracorporeal circuit. Platelets released from artificial surfaces showimpaired hemostatic function. Compounds of the invention can beadministered to prevent adhesion.

[0192] Other applications of these compounds include prevention ofplatelet thrombosis, thromboembolism and reocclusion during and afterthrombolytic therapy and prevention of platelet thrombosis,thromboembolism and reocclusion after angioplasty of coronary and otherarteries and after coronary artery bypass procedures.

[0193] The compounds of the present invention also encompass theirnon-toxic pharmaceutically acceptable salts, such as, but not limitedto, an alkali metal salt such as sodium or potassium; an alkaline earthmetal salt such as manganese, magnesium or calcium; or an ammonium ortetraalkyl ammonium salt, i.e., NX₄ ⁺ (wherein X is C₁₋₄).Pharmaceutically acceptable salts are salts that retain the desiredbiological activity of the parent compound and do not impart undesiredtoxicological effects.

[0194] Those skilled in the art will recognize various syntheticmethodologies which can be employed to prepare non-toxicpharmaceutically acceptable salts and acylated prodrugs of thecompounds.

[0195] The active compounds can be administered systemically to targetsites in a subject in need such that the extracellular concentration ofa P2Y₁₂ agonist is elevated to block the binding of ADP to P2Y₁₂receptor, thus inhibit the platelet aggregation. The term systemic asused herein includes subcutaneous injection, intravenous, intramuscular,intrastemal injection, intravitreal injection, infusion, inhalation,transdermal administration, oral administration, rectal administrationand intra-operative instillation.

[0196] For systemic administration such as injection and infusion, thepharmaceutical formulation is prepared in a sterile medium. The activeingredient, depending on the vehicle and concentration used, can eitherbe suspended or dissolved in the vehicle. Adjuvants such as localanesthetics, preservatives and buffering agents can also be dissolved inthe vehicle. The sterile indictable preparation can be a sterileindictable solution or suspension in a non-toxic acceptable diligent orsolvent. Among the acceptable vehicles and solvents that can be employedare sterile water, saline solution, or Ringer's solution.

[0197] Another method of systemic administration of the active compoundinvolves oral administration, in which pharmaceutical compositionscontaining active compounds are in the form of tablets, lozenges,aqueous or oily suspensions, viscous gels, chewable gums, dispersiblepowders or granules, emulsion, hard or soft capsules, or syrups orelixirs.

[0198] For oral use, an aqueous suspension is prepared by addition ofwater to dispersible powders and granules with a dispersing or wettingagent, suspending agent one or more preservatives, and other excipients.Suspending agents include, for example, sodium carboxymethylcellulose,methylcellulose and sodium alginate. Dispersing or wetting agentsinclude naturally-occurring phosphatides, condensation products of anallylene oxide with fatty acids, condensation products of ethylene oxidewith long chain aliphatic alcohols, condensation products of ethyleneoxide with partial esters from fatty acids and a hexitol, andcondensation products of ethylene oxide with partial esters derived fromfatty acids and hexitol anydrides. Preservatives include, for example,ethyl, and n-propyl p-hydroxybenzoate. Other excipients includesweetening agents (e.g., sucrose, saccharin), flavoring agents andcoloring agents. Those skilled in the art will recognize the manyspecific excipients and wetting agents encompassed by the generaldescription above.

[0199] For oral application, tablets are prepared by mixing the activecompound with nontoxic pharmaceutically acceptable excipients suitablefor the manufacture of tablets. These excipients can be, for example,inert diluents, such as calcium carbonate, sodium carbonate, lactose,calcium phosphate or sodium phosphate; granulating and disintegratingagents, for example, corn starch, or alginic acid; binding agents, forexample, starch, gelatin or acacia; and lubricating agents, for examplemagnesium stearate, stearic acid or talc. The tablets can be uncoated orthey can be coated by known techniques to delay disintegration andabsorption in the gastrointestinal tract and thereby provide a sustainedaction over a longer period. For example, a time delay material such asglyceryl monostearate or glyceryl distearate can be employed.Formulations for oral use can also be presented as hard gelatin capsuleswherein the active ingredient is mixed with an inert solid diluent, forexample, calcium carbonate, calcium phosphate or kaolin, or as softgelatin capsules wherein the active ingredient is mixed with water or anoil medium, for example, peanut oil, liquid paraffin or olive oil.Formulation for oral use can also be presented as chewable gums byembedding the active ingredient in gums so that the active ingredient isslowly released upon chewing.

[0200] Additional means of systemic administration of the activecompound to the target platelets of the subject would involve asuppository form of the active compound, such that a therapeuticallyeffective amount of the compound reaches the target sites via systemicabsorption and circulation.

[0201] For rectal administration, the compositions in the form ofsuppositories can be prepared by mixing the active ingredient with asuitable non-irritating excipient which is solid at ordinarytemperatures but liquid at the rectal temperature and will thereforemelt in the rectum to release the compound. Such excipients includecocoa butter and polyethylene glycols.

[0202] The active compounds can also be systemically administered to theplatelet aggregation sites through absorption by the skin usingtransdermal patches or pads. The active compounds are absorbed into thebloodstream through the skin. Plasma concentration of the activecompounds can be controlled by using patches containing differentconcentrations of active compounds.

[0203] One systemic method involves an aerosol suspension of respirableparticles comprising the active compound, which the subject inhales. Theactive compound would be absorbed into the bloodstream via the lungs,and subsequently contact the target platelets in a pharmaceuticallyeffective amount. The respirable particles can be liquid or solid, witha particle size sufficiently small to pass through the mouth and larynxupon inhalation; in general, particles ranging from about 1 to 10microns, but more preferably 1-5 microns, in size are consideredrespirable.

[0204] Another method of systemically administering the active compoundsto the platelet aggregation sites of the subject involves administeringa liquid/liquid suspension in the form of eye drops or eye wash or nasaldrops of a liquid formulation, or a nasal spray of respirable particlesthat the subject inhales. Liquid pharmaceutical compositions of theactive compound for producing a nasal spray or nasal or eye drops can beprepared by combining the active compound with a suitable vehicle, suchas sterile pyrogen free water or sterile saline by techniques known tothose skilled in the art.

[0205] Intravitreal delivery can include single or multiple intravitrealinjections, or via an implantable intravitreal device that releasesP2Y₁₂ antagonists in a sustained capacity. Intravitreal delivery canalso include delivery during surgical manipulations as either an adjunctto the intraocular irrigation solution or applied directly to thevitreous during the surgical procedure.

[0206] For systemic administration, plasma concentrations of activecompounds delivered can vary according to compounds; but are generally1×10⁻¹⁰−1×10⁻⁵ moles/liter, and preferably 1×10⁻⁸−1×10⁻⁶ moles/liter.

[0207] The pharmaceutical utility of P2Y₁₂ antagonist compounds of thisinvention is indicated by their inhibition of ADP-induced plateletaggregation. This widely used assay, as described in S. M. O. Hourani etal. Br. J. Pharmacol. 105, 453-457 (1992) relies on the measurement ofthe aggregation of a platelet suspension upon the addition of anaggregating agent such as ADP.

[0208] The present invention also provides novel compounds. The presentinvention additionally provides novel pharamaceutical formulationscomprising compounds of Formula I of high purity, and/or in apharmaceutically acceptable carrier. The pharmaceutically acceptablecarrier can be selected by those skilled in the art using conventionalcriteria. The pharmaceutically acceptable carrier include, but are notlimited to, saline and aqueous electrolyte solutions, water polyetherssuch as polyethylene glycol, polyvinyls such as polyvinyl alcohol andpovidone, cellulose derivatives such as methylcellulose andhydroxypropyl methylcellulose, petroleum derivatives such as mineral oiland white petrolatum, animal fats such as lanolin, polymers of acrylicacid such as carboxypolymethylene gel, vegetable fats such as peanut oiland polysaccharides such as dextrans, and glycosaminoglycans such assodium hyaluronate and salts such as sodium chloride and potassiumchloride.

[0209] Novel compounds of the present invention include compounds ofFormula Ib (mononucleotide), provided that when n=1, both X₁ and X₂ arenot 0; and when n=0, X₁ is not 0; and provided when Y′═H, that X₂ isindependently O, CH₂, CHF, CHCl, CF₂, CCl₂; also provided that whenR₁₀═NH₂ or 0, and when R₅ and R₆ are taken together as oxygen doublybonded to C, then R₇ is not equal to ortho-methylamino phenyl; furtherprovided that when n=p=1, X₂═CH₂ and B′=adenosine, then R₁ and R₂ arenot equal to napththylenylmethyl, napthylenylmethylene, orphenylmethylene.

[0210] Novel compounds of the present invention also include compoundsof Formula Ia, wherein B and B′ are independently pyrimidine(pyrimidine/pyrimidine dinucleotide), provided that when m+n+p=1,R₁₆═CH₃, and R₅ and R₆ are taken together as oxygen doubly bonded to C,then R₇ is not equal to CH₃ (Z′ does not equal to acetate); alsoprovided that when m+n+p=3, B and B′=uridine, and R₅ and R⁶ are takentogether as oxygen doubly bonded to C, then R₇ is not equal to phenylfor Y′═OR₁ and/or Y═OR₄ (Y and Y′ does not equal to benzoyl); furtherprovided that when m+n+p=1, then both R₈ and R₉ are not CH₃ (Z′ and Y′taken together do not equal isopropylidine).

[0211] Novel compounds of the present invention also include compoundsof Formula Ia, wherein B is a purine or residue according to generalformula IV, and B′ is a pyrimidine residue according to general formulaV, (purine/pyrimidine dinucleotide); provided that Y′ is not equal toOCH₃ when Z′, Y, or Y′═H or OH; further provided that R₉ is not equal toOCH₂CH₃ when R₉═H(Z′ and Y′ or Z and Y taken together do not equal to anorthoethylester).

[0212] Novel compounds of the present invention also include compoundsof Formula Ia, wherein B and B′ are independently a purine residueaccording to general formula IV, (purine/purine dinonucleotide);provided that (a)Y or Y′ is not equal to OCH₃ when R₁₀═NH₂ or 0; (b)R₈is not equal to OCH₃ or OCH₂CH₃ when R₉═H; (c) both R₈ and R₉ are notequal to CH₃; (d) when m+n+p=1, then R₈ and R₉ does not equal OCH₂CH₃;(e) when R₁₀=NH₂, and when R₅ and R⁶ are taken together as oxygen doublybonded to C, then R₇ is not equal to ortho-methylaminophenyl; (f) whenm+n+p=1, and when R₅ and R₆ are taken together as oxygen doubly bondedto C, then R₇ is not equal to CH(CH₂CH₂SCH₃)NHS(o-NO₂-Ph) orCH(CH₂Ph)NHS(o-NO₂-Ph).

[0213] Preferred compounds of the present invention include 2′- or3′-phenylcarbamate UTP, 2′,3′-di-phenylcarbamate UTP,2′,3′-phenylacetaldehyde acetal ADP, di[3′(phenylcarbamate)dUp2dU],2′,3′-phenylacetaldehyde acetal Up3U, di 2′,3′-phenylacetaldehyde acetalUp3U, 2′,3′-phenylacetaldehyde acetal Up4A, 2′,3′-phenylacetaldehydeacetal Ap4U, di 2′,3′-phenylacetaldehyde acetal Ap4U,2′,3′-phenylacetaldehyde acetal Ip4U, 2′,3′-phenylacetaldehyde acetalUp4U, 2′,3′-phenylacetaldehyde acetal Ip4U, 2′,3′-phenylacetaldehydeacetal Up4dC, tetraphenylcarbamate Up4U, di2′,3′-benzaldehyde acetalIp4U, di 2′,3′-benzaldehyde acetal Up4U, 2′,3′-benzaldehyde acetal Up4U,di 2′,3′-phenylacetaldehyde acetal Cp4U, 2′,3′-phenylacetaldehyde acetalCp4U, 2′,3′-phenylacetaldehyde acetal Up4C, 2′,3′-phenylacetaldehydeacetal Up4T, di 2′,3′-benzaldehyde acetal Cp4U, 2′,3′-benzaldehydeacetal Ip4U, 2′,3′-benzaldehyde acetal Up4U, 2′,3′-benzaldehyde acetalUp4dC, 2′,3′-benzaldehyde acetal Cp4U, 2′,3′-benzaldehyde acetal Up4C,2′,3′-phenylpropionaldehyde acetal Up4U, di 2′,3′-phenylpropionaldehydeacetal Up4U, 2′,3′-benzaldehyde acetal Cp4C, Bis MANT Up4U, Mant Up4U,Di 2′,3′-benzylacetal Up4U, Mono 2′, 3′-benzylacetal Up4U, Triphenylcarbamate Up4U, 2′,3′-phenylcarbamate Up4U, and monophenylcarbamateUp4U.

[0214] Novel mononucleoside 5′-monophosphates compounds includecompounds of Formula Ib′

[0215] wherein:

[0216] V═O;

[0217] A=M;

[0218] M=H or a pharmaceutically-acceptable inorganic or organiccounterion;

[0219] D₁=O;

[0220] Y′═H, OH, or OR₁;

[0221] Z′=H, OH, or OR₂; with the proviso that at least one of Y′ and Z′is OR, or OR₂;

[0222] R₁ and R₂ are residues which are linked directly to the 2′ and/or3′ hydroxyls of the furanose or carbocycle via a carbon atom accordingto Formula II, or linked directly to two of the 2′ and 3′ hydroxyls ofthe furanose or carbocycle via a common carbon atom according to FormulaIII,

[0223] wherein:

[0224] the O atoms are the 2′- and 3′-oxygens of the furanose; and

[0225] the 2′- and 3′-oxygens of the furanose are linked by a commoncarbon atom to form a cyclical acetal; and

[0226] R₈ is hydrogen; and

[0227] R₉ is selected from the group consisting of aralkyl, aryl,substituted aralkyl, and substituted aryl;

[0228] in which the aralkyl groups are straight chained from 1 to 5carbons, with or without unsaturation and without heteroatoms in thealkyl portion, and are monocyclic moieties from 5 to 6 carbons in thearyl portion; and the aryl groups are monocyclic moieties from 4 to 6carbons, with or without heteroatoms;

[0229] B′ is a purine residue according to general Formula IV

[0230] wherein:

[0231] R₁₀ is acylamino, according to Formula VI; and

[0232] R₁₇ is amino or mono- or disubstituted amino such that the moietyaccording to Formula VI is a urea;

[0233] J=carbon;

[0234] R₁₁ is absent;

[0235] R₁₂ is hydrogen; and

[0236] R₁₃ is hydrogen.

[0237] For mononucleoside 5′-monophosphates, preferred compounds of thepresent invention include 2′,3′-phenylacetaldehyde acetal-6-N-phenylureaAMP (compound 22), 2′, 3′-phenylacetaldehyde acetal-6-N-n-hexylurea AMP(compound 23), 2′,3′-phenylacetaldehyde acetal-6-N-ethylurea AMP(compound 24), 2′,3′-phenylacetaldehyde acetal-6-N-cyclopentylurea AMP(compound 25), 2′,3′-cinnamyl acetal-6-N-n-hexylurea AMP (compound 26),2′,3′-cinnamyl acetal-6-N-ethylurea AMP (compound 27), 2′,3′-cinnamylacetal-6-N-phenylurea AMP (compound 28), 2′,3′-cinnamylacetal-6-N-n-propylurea AMP (compound 29), 2′,3′-cinnamylacetal-6-N-n-butylurea AMP (compound 30), 2′,3′-phenylpropargylacetal-6-N-phenylurea AMP (compound 31), 2′,3′-phenylpropargylacetal-6-N-n-hexylurea AMP (compound 32), 2′,3′-phenylpropargylacetal-6-N-n-butylurea AMP (compound 33), 2′,3′-phenylpropargylacetal-6-N-n-propylurea AMP (compound 34), 2′,3′-phenylpropargylacetal-6-N-ethylurea AMP (compound 35), 2′,3′-benzaldehydeacetal-6-N-ethylurea AMP (compound 36), 2′,3′-benzaldehydeacetal-6-N-n-propylurea AMP (compound 37), 2′,3′-benzaldehydeacetal-6-N-n-butylurea AMP (compound 38), 2′,3′-benzaldehydeacetal-6-N-n-hexylurea AMP (compound 39), and 2′,3′-benzaldehydeacetal-6-N-cyclopentylurea AMP (compound 40).

[0238] Preferred compositions also comprise the following Compounds1-40. In the following structures hydrogens which are understood to bepresent have been omitted for the sake of simplicity. Tautomers drawnrepresent all tautomers possible. As diastereomers are generated withthe introduction of the acetal group, structures containing this moietyare taken to mean either of the possible diastereomers alone or amixture of diasteromers in any ratio.

[0239] The invention is illustrated further by the following examplesthat are not to be construed as limiting the invention in scope to thespecific procedures described in them.

EXAMPLES Example 1

[0240] 2′(3′)-O-((phenylaminocarbonyl)-uridine 5′-)triphosphate

[0241] Uridine 5′-triphosphate, ditributylammonium salt (100 mg, 0.176mmol; prepared from the trisodium salt by treatment with Dowex 50W×4H⁺in water, followed by mixing the protonated species with an excess oftributylamine, stripping and lyophilization) was dissolved in dry DMF (1mL) and phenylisocyanate (19 μL, 0.176 mmol) added. The reaction mixturewas heated at 45° C. for 15 minutes, at which point a further portion ofphenylisocyanate (19 μL, 0.176 mmol) was added. The solution was heatedat 45° C. overnight and the DMF was removed on a rotary evaporator. Theresidual oil was partitioned between water (2 mL) and ethyl acetate (2mL) and the layers were separated. The aqueous layer was extracted twicemore with ethyl acetate (2 mL each) and the water was removed on arotary evaporator. The residue was dissolved in water (1.5 mL) and theproduct isolated by repeated injections onto a preparative HPLC column(Alltech Nucleotide/Nucleoside C18, 7 um, 10×250 mm, gradient from 0.1 Mammonium acetate to methanol over 30 minutes, 5 mL/min, monitor at 260nm). The yield of the carbamate was 26 mg (22%, calculated for thetetraammonium salt). 1H NMR showed the product to be a mixture of 2′ and3′ carbamates. The product so obtained can be used for the purposes ofthis invention per se or can be activated with a suitable coupling agent(e.g. a carbodiimide) and reacted with a variety of nucleotides togenerate novel dinucleoside polyphosphates.

[0242] 1H NMR (D20, 300 MHz): δ 4.10-4.47 (m, 4H), 5.17 (m, 1H), 5.83(dd, 1H), 5.96 (m, 1H), 7.04 (t, 1H), 7.25 (m, 4H), 7.79 (m, 1H). ³¹PNMR (D2O,121.47 MHz): 6-9.54 (m, 1P), −10.20 (m, 1P), −21.87 (m, 1P).

Example 2

[0243] 2′(3′)-O-(phenylaminocarbonyl)-P¹,P⁴-di(uridine5′-)tetraphosphate [“monophenylcarbamate Up4U”],Di-2′(3′)-O-(phenylaminocarbonyl)-P¹,P⁴-di(uridine 5′-)tetraphosphate[“diphenylcarbamate Up4U”] andTri-2′(3′)-O-(phenylaminocarbonyl)-P¹,P⁴-di(uridine 5′-)tetraphosphate[“triphenylcarbamate Up4U”]

[0244] P¹,P⁴-Di(uridine 5′-)tetraphosphate, ditributylammonium salt (211mg, 0.182 mmol; prepared from the tetrasodium salt by treatment withDowex 50W×4H⁺ in water, followed by mixing the protonated species withan excess of tributylamine, stripping and lyophilization) was dissolvedin dry DMF (2 mL) and phenylisocyanate (40 μL, 3.64 mmol) added in asingle portion. The homogeneous reaction mixture was heated overnight at45° C., whereupon TLC (silica gel, 50% isopropanol/50% ammoniumhydroxide) indicated a substantial conversion to two products. Thesolvent was removed on a rotary evaporator and the residue waspartitioned between water (7 mL) and ethyl acetate (10 mL). The layerswere separated, and the aqueous was extracted twice more with ethylacetate. (10 mL each). The water was removed from the aqueous extractand the residual oil lyophilized overnight. The solid obtained wasreconstituted in water (3 mL) and the two products separated by repeatedinjections onto a semipreparative HPLC column (AlltechNucleotide/Nucleoside C18, 7 um, 10×250 mm, gradient from 0.1 M ammoniumacetate to methanol over 30 minutes, 5 mL/min, monitor at 260 nm).Stripping and lyophilization gave the mono-phenylcarbamate (48 mg, 27%yield), di-phenylcarbamate (16 mg, 8% yield) and a trace amount of thetriphenylcarbamate, as the tetraammonium salts. All three products weremixtures of the corresponding 2′/3′ regiosiomers.

[0245] Monophenylcarbamate: ¹H NMR (D20, 300 MHz): δ 4.08-4.65 (m, 9H),5.14 (d, 1H), 5.75-5.94 (m, 4H), 7.01 (t, 1H), 7.22 (m, 4H), 7.76 (m,2H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.17 (m, 2P), −21.81 (m, 2P).

[0246] Diphenylcarbamate: ¹H NMR (D₂O, 300 MHz): δ 4.13-4.43 (m, 8H),5.12 (m, 2H), 5.84 (m, 4H), 7.01 (m, 2H), 7.21 (m, 8H), 7.75 (dd, 2H).³¹P NMR (D₂O, 121.47 MHz): 6-10.19 (m, 2P), −21.65 (m, 2P).

[0247] Triphenylcarbamate: ¹H NMR (D₂O, 300 MHz): δ 4.29 (m, 7H), 4.5.10(m, 1H), 5.27 (m, 2H), 5.87 (m, 4H), 7.09 (m, 15H), 7.76 (d, 2H). ³¹PNMR (D₂O, 121.47 MHz): 6-10.30 (m, 2P), −21.73 (m, 2P).

Example 3

[0248] P¹,P⁴-Tetra-(2′(3′)-O-(phenylaminocarbonyl) di(uridine5′-)tetraphosphate [tetraphenylcarbamate Up4U”]

[0249] This derivative was prepared according to the method of example2. P¹,P⁴-Di(uridine 5′-)tetraphosphate, ditributylammonium salt (200 mg,0.172 mmol) was treated with 16 eq of phenylisocyanate (300 uL, 2.76mmol) in DMF and stirred overnight at 35° C. The solvent was evaporatedand the excess reagents removed by extraction of an aqueous solution ofthe product with ethyl acetate. Following preparative HPLC as previouslydescribed, 93 mg (30% yield) of the tetraphenylcarbamate was obtained.

[0250] Tetraphenylcarbamate¹H NMR (D₂O, 300 MHz): δ 7.75 (d, 2H), 7.11(m, 16H), 6.94 (m, 4H), 5.95 (d, 2H), 5.80 (d, 2H), 5.32 (m, 2H), 5.23(m, 2H), 4.42 (m, 2H), 4.25 (m, 2H), 4.16 (m, 2H). ³¹P NMR (D₂O, 121.47MHz):): 6-10.30 (m, 2P), −22.32 (m, 2P).

Example 4

[0251] 2′,3′-(benzyl)methylenedioxy-P¹,P⁴-di(uridine 5′-)tetraphosphate[“Mono 2′/3′ benzylacetal Up4U”] andP¹,P⁴-Di-(2′,3′-((benzyl)methylenedioxy) di(uridine 5′-)tetraphosphate[“Di 2′/3′ benzylacetal Up4U”]

[0252] P¹,P⁴-Di(uridine 5′-)tetraphosphate, tetrasodium salt (290 mg,0.332 mmol) was dissolved in 98% formic acid and phenylacetaldehyde,dimethyl acetal (110 uL, 0.662 mmol) added. The reaction was stirred atambient temperature for 3 days, at which point TLC (silica gel, 50%isopropanol/50% ammonium hydroxide) and HPLC (C 18) showed goodconversion to two less polar products. The formic acid was removed on arotary evaporator, and the residue partitioned between 0.7 M ammoniumbicarbonate (15 mL) and butyl acetate (15 mL). The layers were separatedand the aqueous was washed with a further portion of butyl acetate (10mL). The aqueous layer was stripped and the residue lyophilizedovernight. The crude product was dissolved in water (5 mL) and thecomponents separated by preparative HPLC (Waters Novapak C18, 6 um,25×100 mm, gradient from 0.1 M ammonium acetate to methanol over 30minutes, 30 mL/min, monitor at 260 nm). The yield of the monoacetal was88 mg (28%) and of the diacetal 60 mg (17%), both as the tetraammoniumsalts.

[0253] Monoacetal: ¹H NMR (D₂O, 300 MHz): δ 2.99 (d, 2H), 4.01-4.32 (m,8H), 4.77 (m, 2H), 5.33 (m, 2H), 5.74 (d, 1H), 5.81 (m, 2H), 7.21 (m,5H), 7.64 (d, 1H), 7.79 (d, 1H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.18 (m,1P), −10.78 (m, 1P), −22.00 (m, 2P).

[0254] Diacetal: ¹H NMR (D₂O, 300 MHz): δ 2.98 (d, 4H), 3.99 (m, 4H),4.27 (m, 2H), 5.27 (m, 2H), 5.36 (m, 2H), 5.73 (d, J=8.1 Hz, 2H), 7.21(m, 10H), 7.61 (d, J=8.1 Hz, 2H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.57 (m,2P), −21.81 (m, 2P).

Example 5

[0255] 2′,3′-((benzyl)methylenedioxy) P¹,P³-uridine 5′-)triphosphate[“2′3′ phenylacetaldehyde acetal Up3U”] andP¹,P³-Di-(2′,3′-((benzyl)methylenedioxy) uridine 5′-)triphosphate [“di2′3′ phenylacetaldehyde acetal Up3U”]

[0256] P¹,P³-Di(uridine 5′-)triphosphate, trisodium salt (100 mg, 0.129mmol) was dissolved in 98% formic acid and phenylacetaldehyde, dimethylacetal (64 uL, 0.386 mmol) added. After overnight stirring at roomtemperature, the formic acid was removed, and the residue partitionedbetween 1 M sodium bicarbonate and ethyl acetate. Following removal ofthe organic layer, the product was purified on preparative HPLC, aspreviously described. Following lyophilization, 40 mg (36%) of themonoacetal and 24 mg (19%) of the diacetal were obtained.

[0257] Monoacetal: ¹H NMR (D₂O, 300 MHz): δ 7.7s (d, 2H), 7.54 (d, 2H),7.16 (s, 5H), 5.70 (m, 3H), 5.31 (s, 1H), 5.23 (s, 1H), 4.66 (m, 2H),4.10 (m, 8H), 2.93 (d, 2H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.30 (m, 1P),10.81 (m, 1P), −21.99 (m, 1P).

[0258] Diacetal: ¹H NMR (D₂O, 300 MHz): δ 7.51 (d, 2H), 7.15 (m, 10H),5.65 (d, 2H), 5.31 (d, 2H), 5.20 (t, 2H), 4.63 (m, 2H), 4.13 (m, 2H),3.88 (m, 4H), 2.90 (d, 4H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.75 (m, 2P),−21.97 (m, 1P).

Example 6

[0259] P¹-2′,3′-((benzyl)methylenedioxy) (uridine 5′-) P⁴-(deoxycytidine5′-)tetraphosphate [“2′3′ phenylacetadehyde acetal Up4dC”]

[0260] P¹-(uridine 5′-) P⁴-(deoxycytidine 5′-)tetraphosphate,tetrasodium salt (100 mg, 0.16 mmol) was dissolved in 98% formic acid (1mL), and phenylacetaldehyde, dimethyl acetal (57 uL, 0.384 mmol) added.After overnight stirring, the formic acid was removed and the residuepartitioned between 1 M sodium bicarbonate and ethyl acetate. Afterseparation of the layers, the product was purified on preparative HPLC,as previously described. Yield 40 mg (36%). This product was amenable tosubsequent modification of the deoxy cytidine base by the proceduresdescribed in examples 9-13, giving rise to lipophilic bifunctionalmolecules falling within the scope of this invention.

[0261] Monoacetal: ¹H NMR (D₂O, 300 MHz): δ 7.98 (d, 1H), 7.62 (d, 1H),7.21 (m, 5H), 6.11 (m, 2H), 5.74 (d, 1H), 5.39 (d, 1H), 5.31 (t, 1H),4.77 (m, 2H), 4.45 (m, 1H), 4.32 (m, 1H), 4.03 (m, 5H), 2.99 (d, 2H),2.29 and 2.21 (M, 2H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.15 (m, 1P),−10.68 (m, 1P), −21.98 (m, 2P).

Example 7

[0262] 3′-O-(phenylaminocarbonyl)-2′-deoxy(uridine 5′)-monophosphate

[0263] Deoxyuridine 5′-monophosphate, tetrabutylammonium salt (135 mg,0.274 mmol; prepared from the disodium salt by treatment with Dowex50W×4H⁺, followed by stirring the resultant neutral species with excesstributylamine, stripping and lyophilization) was dissolved in dry DMF (1mL). Phenylisocyanate (60 uL, 0.547 mmol) was added and the mixtureheated overnight at 45° C., at which time TLC (silica gel, 50%isopropanol/50% ammonium hydroxide) and HPLC (C 18) indicated asubstantial conversion to a less polar product. The DMF was stripped ona rotary evaporator and the oily residue partitioned between water (10mL) and ethyl acetate (10 mL). The layers were separated and the aqueouslayer was rewashed with ethyl acetate (2×10 mL). The water was removedand the residue was dissolved in water (2 mL). The product was isolatedby repeated injections onto semipreparative HPLC (AlltechNucleotide/Nucleoside C18, 7 um, 10×250 mm, gradient from 0.1 M ammoniumacetate to methanol over 30 minutes, 5 mL/min, monitor at 260 nm). Theyield was 67 mg as the diammonium salt (53%).

[0264]¹H NMR (D₂O, 300 MHz): δ 2.21 (m, 2H), 3.84 (s, 2H), 4.13 (s, 1H),5.08 (d, 1H), 5.63 (d, 1H), 6.06 (t, 1H), 6.89 (br. t, 1H), 7.10 (m,4H), 7.72 (d, 1H).

[0265]³¹P NMR (D₂O, 121.47 MHz): δ−2.31 (s).

[0266] P¹-(3′-O-(phenylaminocarbonyl)-2′-deoxyuridine 5′-)P⁴-(uridine5′-)tetraphosphate

[0267] Uridine 5′-triphosphate, ditributylammonium salt (prepared fromthe trisodium salt by treatment with Dowex 50W×4H⁺, followed by stirringthe resultant neutral species with excess tributylamine, stripping andlyophilization) is treated with 1.5 equivalents ofdicyclohexylcarbodiimide in DMF for 2 hours at room temperature. Thedicyclohexylurea is filtered off, and the resultant uridine 5′-cyclicaltriphosphate is treated with 3′-O-(phenylaminocarbonyl)-2′-deoxy(uridine5′)-monophosphate, which is in the monotributylammonium salt form. Thereaction mixture is stirred for several days at 45° C., and the solventis removed. The products are separated by preparative HPLC, as has beenpreviously described.

Example 8

[0268] 2′(3′)-(2-methylamino)benzoyl-P¹,P⁴-di(uridine 5′-)tetraphosphate(“MANT Up4U”) and P¹,P⁴-Di-(2′(3′)-(2-methylamino)benzoyl uridine5′-)tetraphosphate (“Bis MANT Up4U”)

[0269] P¹,P⁴-Di(uridine 5′-)tetraphosphate, tetrasodium salt (800 mg,0.93 mmol) was dissolved in water (5 mL) and the pH adjusted to 7.6 bythe addition of solid sodium bicarbonate. N,N-dimethylformamide(DMF, 5mL) was added, followed by N-methylisatoic anhydride (231 mg, 1.3 mmol)and the suspension was heated at 50° C. for 2.5 hrs. TLC (silica gel,50% isopropanol, 50% ammonium hydroxide) indicated that the reaction wasnot done by this time, so a further portion of N-methylisatoic anhydride(100 mg, 0.56 mmol) was added and the reaction heated for another hour.The DMF was removed on a rotary evaporator and the residue was dissolvedin a minimum of water and applied to a DEAE Sephadex A-25 column (3×60cm). The column was eluted with a stepwise gradient from water to 1 Mammonium bicarbonate and the eluent monitored with a UV detector set at254 nm. The two products that eluted were collected separately and thesolvent was removed from each and the residue lyophilized overnight. ¹HNMR indicated that the first product to elute was the monoacylatedcompound, while the latter was the diacylated derivative, and that bothwere mixtures with the acylation at either the 2′ or 3′ hydroxyls, butwithout two carbamates on the same sugar. The yield of themonoaminobenzoylated product was 150 mg (16%); the yield of thediaminobenzoylated compound was 91 mg (8.7%).

[0270] Monoaminobenzoylated derivative: ¹H NMR (D₂O, 300 MHz): δ 2.70(s, 3H), 4.09-4.55(m, 9H), 5.34 (m, 1H), 5.71 (m, 2H), 5.83 (dd, 1H),6.01 (m, 1H), 6.57 (m, 1H), 6.65 (m, 1H), 7.25 (t, 1H), 7.72 (d, 2H),7.81 (m, 2H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.20 (m, 2P), −21.83 (m,2P).

[0271] Diaminobenzoylated derivative: ¹H NMR (D₂O, 300 MHz): δ 2.69 (s,6H), 4.15-4.51 (m, 8H), 5.27 (m, 2H), 5.86 (m, 4H), 6.60 (m, 4H), 7.30(m, 2H), 7.79 (m, 4H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.16 (m, 2P),−21.76 (m, 2P).

Example 9

[0272] P¹-(4-N-(4-methoxyphenyl)aminocarbonylcytidine 5′-)-P⁴-(uridine5′-)tetraphosphate

[0273] P¹-(cytidine 5′-)-P⁴-(uridine 5′-)tetraphosphate,ditributylammonium salt (50 mg, 0.043 mmol; prepared from thetetraammonium salt by treatment with Dowex 50W×4H⁺ in water, followed bymixing the protonated species with an excess of tributylamine inmethanol, stripping and lyophilization) was dissolved in dry DMF (1 mL)and tributylamine (10 uL, 0.43 mmol), and p-methoxyphenylisocyanate (8.4uL, 0.648 mmol) were added in a single portion. The homogeneous reactionmixture was heated overnight at 35° C., whereupon TLC (silica gel, 50%isopropanol/50% ammonium hydroxide) and HPLC (C18) indicated asubstantial conversion to a single product. The solvent was removed on arotary evaporator and the residue dissolved in water (1 mL). The productwas isolated by repeated injections onto a semi-preparative HPLC column(Alltech Nucleotide/Nucleoside C18, 7 um, 10×250 mm, gradient from 0.1 Mammonium acetate to methanol over 30 minutes, 5 mL/min, monitor at 260nm). Stripping and lyophilization gave the p-methoxyphenylurea (24 mg,55% yield), as the tetraammonium salt.

[0274] The product so obtained can be derivatized on the 2′ and/or 3′hydroxyl groups according to the foregoing methods (e.g. Examples 2-6).

[0275]¹H NMR (D₂O, 300 MHz): δ 3.59 (s, 3H), 4.01-4.20 (m, 10H), 5.68(m, 3H), 6.19 (d, 1H), 6.71 (d, 2H), 7.18 (d, 2H), 7.67 (d, 1H), 8.06(d, 1H). ³¹P NMR (D₂O, 121.47 MHz): 6-10.13 (m, 2P), −21.76 (m, 2P).

Example 10

[0276] P¹-((4-bromophenyl)ethenocytidine 5′-)-P⁴-(uridine5′-)tetraphosphate

[0277] P¹-(cytidine 5′-)-P⁴-(uridine 5′-)tetraphosphate, tetrasodiumsalt (500 mg, 0.57 mmol) was dissolved in water (5 mL) and a solution of2,4′-dibromoacetophenone (792 mg, 2.85 mmol) in DMF (15 mL) added. Themixture was heated overnight at 40° C., and a further portion of thedibromoketone (400 mg, 1.44 mmol) in DMF (5 mL) added. The rection washeated a further 5 hrs, and the solvents removed by evaporation. Theresidue was partitioned between water (20 mL) and ethyl acetate (25 mL)and the layers separated. The aqueous layer was washed with furtherethyl acetate (2×15 mL) and the aqueous evaporated to dryness. Theresidue was dissolved in water (5 mL) and the product was isolated byrepeated injections onto a semi-preparative HPLC column (see example 6for conditions). The yield of the pure etheno compound was 80 mg (13.5%)

[0278]¹H NMR (D₂O, 300 MHz): δ 4.06 (m, 8H), 4.36 (m, 2H), 5.64 (dd,2H), 6.07 (d, 1H), 6.74 (d, 1H), 7.45 (d, 2H), 7.54 (d, 2H), 7.59 (d,1H), 7.63 (d, 1H), 7.93 (s, 1H). ³¹P NMR (D₂O, 121.47 MHz): 8-10.09 (m,2P), −21.59 (m, 2P).

Example 11

[0279] P¹-((4-bromophenyl)etheno-2′-deoxycytidine 5′-)-P⁴-(uridine5′-)tetraphosphate

[0280] Example 11 product was prepared from 100 mg P¹-(2′-deoxycytidine5′-)-P⁴-(uridine 5′-)tetraphosphate, tetrasodium salt and2,4′-dibromoacetophenone, according to the general method of example 10.Yield=35 mg (30%).

[0281]¹H NMR (D₂O, 300 MHz): δ 2.31 (m, 2H), 4.03 (m, 8H), 5.60 (dd,2H), 6.41 (t, 1H), 6.73 (d, 1H), 7.53 (m, 5H), 7.65 (d, 1H), 7.93 (s,1H). ³¹P NMR (D₂O, 121.47 MHz): 8-10.11 (m, 2P), −21.58 (m, 2P).

Example 12

[0282] P¹, P⁴-Di((4-bromophenyl)ethenocytidine 5′-)-tetraphosphate

[0283] Example 12 product was prepared from 50 mg P¹,P⁴-Di(cytidine5′-)tetraphosphate, tetrasodium salt and 2,4′-dibromoacetophenone,according to the general method of example 10. Yield=20 mg (29%).

[0284]¹H NMR (D20, 300 MHz): δ 4.24 (m, 10H), 5.98 (d, 2H), 6.39 (d,2H), 7.14 (m, 8H), 7.45 (m, 4H).). ³¹P NMR (D₂O, 121.47 MHz): 8-10.13(m, 2P), −21.68 (m, 2P).

Example 13

[0285] P¹-((4-phenylphenyl)ethenocytidine 5′-)-P⁴-(cytidine5′-)tetraphosphate

[0286] Example 13 product was prepared from 50 mg P¹,P⁴-Di(cytidine5′-)tetraphosphate, tetrasodium salt and 2-bromo-4′-phenylacetophenone,according to the general method of example 10. Yield=15 mg (13%).

[0287]¹H NMR (D20, 300 MHz): δ 4.10 (m, 1OH), 5.48 (d, 1H), 5.87 (m,2H), 6.68 (d, 1H), 7.20 (m, 3H), 7.36 (m, 6H), 7.68 (m, 3H). ³¹P NMR(D₂O, 121.47 MHz): 6-10.08 (m, 2P), −21.78 (m, 2P).

[0288] The products of examples 11-13 can be further derivatizedaccording to the methods of Examples 1-8, to give bifunctional moleculesthat fall within the scope of the invention.

Example 14

[0289] 2′,3′-phenylacetaldehyde acetal adenosine 5′-monophosphate

[0290] Adenosine 5′-monophosphate, free acid (10.0 g, 28.8 mmol) wasdissolved in trifluoroacetic acid (50 mL) and phenylacetaldehyde,dimethylacetal (18.50 mL, 121 mmol) added. The reaction was stirred atambient temperature for 3 hours, after which the trifluoroacetic acidwas evaporated and the residue partitioned between 1 M sodiumbicarbonate (80 mL) and ethyl acetate (40 mL). The layers wereseparated, and the product was isolated from the aqueous layer via C₁₈preparative HPLC. Yield=7.50 g (59%). The ammonium salt so obtained wasconverted to the mono-tributylammonium salt via treatment with a slightexcess of tributylamine in aqueous N,N-dimethylformamide, followed byevaporation and drying.

[0291]¹H NMR (D₂O, 300 MHz): δ 3.06 (d, 2H), 3.86 (m, 2H), 4.39 (m, 1H),4.91 (m, 1H), 5.18 (m, 1H), 5.36 (t, 1H), 5.63 (d, 1H), 7.23 (m, 5H),8.09 (s, 1H), 8.20 (s, 1H). ³¹P NMR (D₂O, 121.47 MHz): δ 2.17 (s).

Example 15

[0292] 2′,3′-cinnamyl acetal adenosine 5′-monophosphate

[0293] Adenosine 5′-monophosphate, free acid (1.0 g, 2.88 mmol) wasdissolved in 98% formic acid (5 mL) and cinnamaldehyde (1.14 g, 8.65mmol) added. The reaction was stirred at ambient temperature for 3hours, after which the formic acid was evaporated and the residuepartitioned between 1 M sodium bicarbonate (25 mL) and ethyl acetate (20mL). The layers were separated, and the product was isolated from theaqueous layer via preparative HPLC. Yield=0.202 g (15%).

[0294]¹H NMR (D20, 300 MHz): δ 3.97 (m, 2H), 4.50 (m, 1H), 5.04 (m, 1H),5.29 (m, 1H), 5.65 (d, 0.4H), 5.86 (d, 0.6H), 6.24 (m, 2H), 6.87 (dd,1H), 7.27 (m, 3H), 7.43 (m, 2H), 8.12 (d, 1H), 8.28 (d, 1H). ³¹P NMR(D₂O, 121.47 MHz): δ 1.42 (d).

Example 16

[0295] 2′,3′-phenylacetaldehyde acetal-6-N-phenylurea adenosine5′-monophosphate (compound 22)

[0296] 2′,3′-phenylacetaldehyde acetal adenosine 5′-monophosphate,tributylammonium salt (prepared according to example 14, 1.0 g, 2.15mmol) was dissolved in N,N-dimethylformamide (10 mL) andphenylisocyanate (1.17 g, 10.72 mmol) added. The reaction was heated at35° C. for 4 hrs, after which the solvent was removed and the residuepartitioned between 1 M sodium bicarbonate (30 mL) and ethyl acetate (25mL). The layers were separated and the product isolated from the aqueouslayer via preparative HPLC. Yield=0.85 g (68%).

[0297]¹H NMR (D₂O, 300 MHz): δ 2.97 (d, 2H), 3.81 (m, 2H), 4.31 (m, 1H),4.78 (m, 1H), 4.98 (m, 1H), 5.23 (t, 1H), 5.63 (d, 1H), 6.74 (m, 1H),6.96 (m, 4H), 7.19 (m, 5), 8.12 (s, 1H), 8.30 (s, 1H). ³¹P NMR (D₂O,121.47 MHz): δ 1.19 (s).

Example 17

[0298] 2′,3′-cinnamyl acetal-6N-ethylurea adenosine 5′-monophosphate(compound 27)

[0299] Compound 27 was prepared according to example 16, starting with2′3′ cinnamyl acetal adenosine 5′-monophosphate (example 15) andsubstituting ethyl isocyanate for phenylisocyanate. Yield=65%.

[0300] 1H NMR (D₂O, 300 MHz): δ 1.07 (t, 3H), 3.21 (q, 2H), 3.93 (m,2H), 4.45 (m, 1H), 4.99 (m, 1H), 5.28 (m, 1H), 5.54 (d, 0.3H), 5.70 (d,0.7H), 5.95 (m, 1H), 6.14 (m, 1H), 6.61 (dd, 1H), 7.14 (m, 5H), 8.29 (m,2H). ³¹P NMR (D₂O, 121.47 MHz): δ 1.93 (d).

Example 18

[0301] Inhibition of ADP-Induced Platelet Aggregation

[0302] Isolation of platelets: Human blood was obtained from informedhealthy volunteers. Blood was collected into one-sixth volume of ACD(2.5 g of sodium citrate, 1.5 g citric acid, and 2.5 g glucose in 100 mldH₂O). Blood was centrifuged at 800×g for 15 min at room temperature andthe platelet-rich plasma removed and incubated for 60 min at 37° C. inthe presence of 1 mM acetylsalicylic acid followed by centrifugation at1000×g for 10 min at room temperature. The platelet pellet wasresuspended at a density of 2×1 8 cells/ml with HEPES-buffered Tyrode'ssolution (137 mM NaCl, 2.7 mM KCl, 1 mM MgCl₂, 3 mM NaH₂PO₄, 5 mMglucose, 10 mM HEPES pH 7.4, 0.2% bovine serum albumin, and 0.05 U/mlapyrase).

[0303] Aggregation Studies: ADP-induced platelet aggregation wasdetermined by measuring the transmission of light through a 0.5 mlsuspension of stirred (900 rpm) aspirin-treated washed platelets in alumi-aggregometer at 37° C. (Chrono-Log Corp. Havertown, Pa.). Thebaseline of the instrument was set using 0.5 ml of Hepes-bufferedTyrode's solution. Prior to aggregation measurements, the plateletsuspension was supplemented with 2 mM CaCl₂ and 1 mg/ml fibrinogen.Platelet aggregation was initiated by the addition of indicatedconcentrations of ADP or other agonists, and the light transmissioncontinuously recorded for at least 8 min. When inhibitors of plateletaggregation were tested, platelets were incubated for 3-6 min in thepresence of indicated concentrations of inhibitor before addition of ADPor other agonists, and the response recorded for at least 8 min. Thepotency of agonists and inhibitors of platelet aggregation wascalculated from both, the rate of aggregation and the maximal extent ofaggregation obtained for each determination by fitting the data to afour-parameter logistic equation using the GraphPad software package(GraphPad Corp. San Diego, Calif.).

[0304] The ability of P2Y₁₂ antagonists to inhibit platelet aggregationis presented in this application as the percent inhibition of theaggregation induced by a maximally effective concentration of ADP. Whena broad range of concentrations of P2Yl₂ antagonist was tested (usuallyfrom 1 nM to 100 μM), an IC₅₀ value was also obtained. IC₅₀ valuesrepresent the concentration of antagonist needed to inhibit by 50% theaggregation elicited by a given concentration of ADP.

Example 19

[0305] Effects on Platelet Aggregation In Vivo

[0306] To evaluate the ability of these compounds to inhibit plateletaggregation in vivo, an experimental protocol similar to the method ofR. G. Humphries et al. (Br. J. Pharmacol. 115:1110-1116, 1995) will beperformed.

[0307] Surgical Preparation and Instrumentation: Male Sprague-Dawleyrats are anesthetized. Body temperature is maintained at 37±0.5° C. witha heating lamp. Animals breathe spontaneously and a tracheotomy isperformed to ensure a patent airway. A cannula containing heparinizedsaline is introduced into the left femoral artery and connected to atransducer to record blood pressure and heart rate. Cannulae containingnon-heparinized saline are introduced into the left common carotidartery and left jugular vein for withdrawal of arterial blood samplesand i.v. administration of compounds, respectively. ExperimentalProtocol: Either compound or vehicle is administered to each animal asan infusion. Blood samples are taken immediately prior to the firstinfusion, at the end of each infusion and 20 min after cessation of thefinal infusion for measurement of platelet aggregation ex vivo.Immediately after sampling, ADP-induced platelet aggregation is measuredin duplicate in 0.5 ml blood samples diluted 1:1 with saline andincubated at 37° C. for 4 min. For the final minute of this period,cuvettes are transferred to a lumi-aggregometer and the sample stirredat 900 rpm. ADP (3 μM) is added in a volume of 20 μl and the aggregationresponse is recorded.

Example 20

[0308] Inhibition of Thrombus Formation in Anesthetized Rats

[0309] To evaluate the effect of these compounds on thrombus formationin vivo, the following experimental protocol will be performed.

[0310] Rats (CD-1; male; approximately 350 grams; Charles River,Raleigh, N.C.), are anesthetized with sodium pentobarbital (70 mg/kgi.p.). The abdomens are shaved and a 22 gauge intravenous catheter isinserted into a lateral tail vein. A midline incision is made and theintestines are wrapped in saline-soaked gauze and positioned so theabdominal aorta is accessible. The inferior vena cava and abdominalaorta are carefully isolated and a section (approx. 1 cm) of theabdominal aorta (distal to the renal arteries proximal to thebifurcation) is dissected. All branches from the aorta in this sectionare ligated with 4-0 silk suture. A 2.5 mm diameter flow probe connectedto a Transonic flow meter is placed on the artery and a baseline(pre-stenosis) flow is recorded. Two clips are placed around the arterydecreasing the vessel diameter by approximately 80%. A second baselineflow measurement is taken (post-stenosis) and the hyperemic response istested. Animals are then treated with either compound or saline i.v.,via tail vein catheter. Thrombosis is induced five minutes aftertreatment by repeated external compressions of the vessel withhemostatic forceps. Two minutes post-injury, the vessel compressions arerepeated and a 10 minute period of flow monitoring is started. Animalsare monitored continuously for a minimum of the first ten minutespost-injury. After twenty minutes (post-injury), a flow measurement isrepeated and the animals are euthanized. The section of the aorta thatincludes the injured section is harvested and placed in 10% formalin forpossible histologic evaluation.

Example 21

[0311] Inhibition of Thrombus Formation in Anesthetized Dogs

[0312] To evaluate the effect of these compounds on dynamic thrombusformation in vivo, the following experimental protocol similar to themethod of J. L. Romson et al. (Thromb. Res. 17:841-853, 1980) will beperformed.

[0313] Surgical Preparation and Instrumentation: Briefly, purpose-breddogs are anesthetized, intubated and ventilated with room air. The heartis exposed by a left thoracotomy in the fifth intercostal space andsuspended in a pericardial cradle. A 2-3 cm segment of the leftcircumflex coronary artery (LCCA) is isolated by blunt dissection. Theartery is instrumented from proximal to distal with a flow probe, astimulation electrode, and a Goldblatt clamp. The flow probe monitorsthe mean and phasic LCCA blood flow velocities. The stimulationelectrode and its placement in the LCCA and the methodology to induce anocclusive coronary thrombus have been described previously (J. K.Mickelson et al., Circulation 81:617-627, 1990; R. J. Shebuski et al.,Circulation 82:169-177, 1990; J. F. Tschopp et al., Coron. Artery Dis.4:809-817, 1993).

[0314] Experimental Protocol: Dogs are randomized to one of fourtreatment protocols (n=6 per treatment group) in which the control groupreceives saline i.v. and the three drug-treated groups are administeredcompound i.v. Upon stabilization from the surgical interventions, dogsreceive either saline or compound. After approximately 30 minutes, ananodal current is applied to the LCCA for 180 min. The number andfrequency of cyclic flow variations (CFV) that precede formation of anocclusive thrombus are recorded. These cyclic phenomena are caused byplatelet thrombi that form in the narrowed lumen as a result of plateletaggregation (J. D. Folts et al., Circulation 54:365-370, 1976; Bush etal., Circulation 69:1161-1170, 1984). Zero flow in the LCCA for aminimum of 30 minutes indicates a lack of antithrombotic efficacy (L. G.Frederick et al., Circulation 93:129-134, 1996).

Example 22

[0315] ADP-Induced Aggregation of Different Compounds

[0316] Different compounds were tested for their inhibition ofADP-induced aggregation and their IC₅₀ according to the protocols inExample 18; the results are shown in FIG. 1. The bar graphs in thefigure illustrate the effect of 100 μM concentration of the compound onADP-induced platelet aggregation, and the data are expressed as %inhibition of the ADP response.

[0317]FIG. 1 shows the structure and abbreviated name of each compoundand its activity. Where hydrogens are understood to be present, theyhave been omitted for the sake of simplicity. For example, for the firststructure of the figure, it is implied that there are hydrogens at the3-position of the pyrimidine ring, at the 3′-position of the ribose onthe oxygen, and on the nitrogen of the carbamate at the 2′-position ofthe ribose. In addition, as disclosed within the scope of the presentinvention, it is implied that the oxygens that are not doubly bonded tothe phosphorous atoms are either present in the ionized form as saltswith a counterion, or are bonded to a hydrogen atom. For simplicity,some of the structures in the figure are portrayed in the salt form, butthis should not be interpreted as excluding the possibility thathydrogens could be present instead.

[0318] Several parent compounds, Up4U, Ip4U, Up3U, and Cp4U, withoutmodifications on the furanose hydroxyl groups, have been included at theend of the figure to illustrate the utility of the present invention.However, these unmodified parent compounds do not inhibit theADP-induced aggregation and are not within the scope of the presentinvention.

[0319] Although the invention has been described with reference to thepresently preferred embodiments, it should be understood that variousmodifications could be made without departing from the scope of theinvention.

1. A compound of Formula Ib′:

wherein: V═0; A=M; M=H or a pharmaceutically-acceptable inorganic ororganic counterion; D₁=O; Y′=OR₁; Z′=OR₂; R₁ and R₂ are residues whichare linked directly to two of the 2′ and 3′ hydroxyls of the furanosevia a common carbon atom according to Formula III,

wherein: O is the 2′ and 3′ oxygens of the furanose; and the 2′ and 3′oxygens of the furanose are linked by a common carbon atom to form acyclical R₈ is hydrogen; and R₉ is selected from the group consisting ofaralkyl, aryl, substituted aralkyl, and substituted aryl; in which thearalkyl groups are straight chained from 1 to 5 carbons, with or withoutunsaturation and without heteroatoms in the alkyl portion, and aremonocyclic moieties from 5 to 6 carbons in the aryl portion; and inwhich the aryl groups are monocyclic moieties from 4 to 6 carbons, withor without heteroatoms; B′ is a purine residue according to generalFormula IV:

wherein J=carbon; R₁₁ is absent; R₁₂ is hydrogen; R₁₃ is hydrogen; R¹⁰is acylamino, according to Formula VI;

wherein: NH is the amino residue at the C-6 position in a purine; C is acarbon atom; W¹ is oxygen; and R₁₇ is amino or mono- or disubstitutedamino such that the moiety according to Formula VI is a urea.
 2. Thecompound according to claim 1, wherein R₉ is selected from the groupconsisting of aralkyl, aryl, substituted aralkyl, and substituted aryl;in which the aralkyl groups are straight chained from 3 to 4 carbons,with or without unsaturation and without heteroatoms in the alkylportion, and are monocyclic moieties of 6 carbons without heteroatoms inthe aryl portion; and the aryl groups are monocyclic moieties of 6carbons, without heteroatoms.
 3. A compound selected from the groupconsisting of: 2′3′ phenylacetaldehyde acetal-6-N-phenylurea AMP; 2′3′phenylacetaldehyde acetal-6-N-n-hexylurea AMP; 2′3′ phenylacetaldehydeacetal-6-N-ethylurea AMP; 2′3′ phenylacetaldehydeacetal-6-N-cyclopentylurea AMP; 2′3′ cinnamyl acetal-6-N-n-hexylureaAMP; 2′3′ cinnamyl acetal-6-N-ethylurea AMP; 2′3′ cinnamylacetal-6-N-phenylurea AMP; 2′3′ cinnamyl acetal-6-N-n-propylurea AMP;2′3′ cinnamyl acetal-6-N-n-butylurea AMP; 2′3′ phenylpropargylacetal-6-N-phenylurea AMP; 2′3′ phenylpropargyl acetal-6-N-n-hexylureaAMP; 2′3′ phenylpropargyl acetal-6-N-n-butylurea AMP; 2′3′phenylpropargyl acetal-6-N-n-propylurea AMP; 2′3′ phenylpropargylacetal-6-N-ethylurea AMP; 2′3′ benzaldehyde acetal-6-N-ethylurea AMP;2′3′ benzaldehyde acetal-6-N-n-propylurea AMP; 2′3′ benzaldehydeacetal-6-N-n-butylurea AMP; 2′3′ benzaldehyde acetal-6-N-n-hexylureaAMP; and 2′3′ benzaldehyde acetal-6-N-cyclopentylurea AMP.
 4. Apharmaceutical formulation comprising the compound according to claim 1and a pharceutically acceptably pharmaceutically acceptable carrier. 5.A pharmaceutical formulation comprising the compound according to claim3 and a pharceutically acceptably pharmaceutically acceptable carrier.6. A method of preventing or treating diseases or conditions associatedwith platelet aggregation comprising: administering to a subject thepharmaceutical formulation according to claim 4, wherein said compoundis effective to bind P2Y₁₂ receptors on platelets and inhibitADP-induced platelet aggregation.
 7. A method of preventing or treatingdiseases or conditions associated with platelet aggregation comprising:administering to a subject the pharmaceutical formulation according toclaim 5, wherein said compound is effective to bind P2Y₁₂ receptors onplatelets and inhibit ADP-induced platelet aggregation.
 8. The methodaccording to claim 6, wherein said pharmaceutical composition reducesthe incidence of dose-related adverse side effects of other therapeuticagents that are used to prevent, manage or treat platelet aggregationdisorders.
 9. The method according to claim 6, wherein said diseases orconditions associated with platelet aggregation are disorders orprocedures characterized by thrombosis, primary arterial thromboticcomplications of atherosclerotic disease, thrombotic complications ofinterventions of atherosclerotic disease, thrombotic complications ofsurgical or mechanical damage, mechanically-induced platelet activation,shunt occlusion, thrombosis secondary to vascular damage andinflammation, indications with a diffuse thrombotic/platelet consumptioncomponent, venous thrombosis, coronary arterial thrombosis, pathologicaleffects of atherosclerosis and arteriosclerosis, platelet aggregationand clot formation in blood and blood products during storage, chronicor acute states of hyper-aggregability, reocclusion of an artery or veinfollowing fibrinolytic therapy, platelet adhesion associated withextracorporeal circulation, thrombotic complications associated withthrombolytic therapy, thrombotic complications associated with coronaryand other angioplasty, or thrombotic complications associated withcoronary artery bypass procedures.
 10. The method according to claim 9,wherein said disorders or procedures associated with thrombosis areunstable angina, coronary angioplasty, or myocardial infarction; saidprimary arterial thrombotic complications of atherosclerosis arethrombotic stroke, peripheral vascular disease, or myocardial infarctionwithout thrombolysis; said thrombotic complications of interventions ofatherosclerotic disease are angioplasty, endarterectomy, stentplacement, coronary or other vascular graft surgery; said thromboticcomplications of surgical or mechanical damage are associated withtissue salvage following surgical or accidental trauma, reconstructivesurgery including skin flaps, or reductive surgery; saidmechanically-induced platelet activation is caused by cardiopulmonarybypass resulting in microthromboembolism and storage of blood products;said shunt occlusion is renal dialysis and plasmapheresis; saidthromboses secondary to vascular damage and inflammation are vasculitis,arteritis, glomerulonephritis or organ graft rejection; said indicationswith a diffuse thrombotic/platelet consumption component aredisseminated intravascular coagulation, thrombotic thrombocytopenicpurpura, hemolytic uremic syndrome, heparin-induced thrombocytopenia, orpre-eclampsia/eclampsia; said venous thrombosis are deep veinthrombosis, veno-occlusive disease, hematological conditions, ormigraine; and said coronary arterial thrombosis is associated withunstable angina, coronary angioplasty or acute myocardial infarction.11. The method according to claim 10, wherein said hematologicalconditions are thrombocythemia or polycythemia.
 12. The method accordingto claim 9, wherein said pathological effects of atherosclerosis andarteriosclerosis are arteriosclerosis, acute myocardial infarction,chronic stable angina, unstable angina, transient ischemic attacks,strokes, peripheral vascular disease, arterial thrombosis, preeclampsia,embolism, restenosis or abrupt closure following angioplasty, carotidendarterectomy, or anastomosis of vascular grafts; said chronic or acutestates of hyper-aggregability is caused by DIC, septicemia, surgical orinfectious shock, post-operative trauma, post-partum trauma,cardiopulmonary bypass surgery, incompatible blood transfusion, abruptioplacenta, thrombotic thrombocytopenic purpura, snake venom or immunediseases.
 13. The method according to claim 9, wherein said reocclusionof an artery or vein following fibrinolytic therapy is inhibited byinternal administration of said compound with a fibrinolytic agent. 14.The method according to claim 13, wherein said fibrinolytic agent is anatural or synthetic product which directly or indirectly causes lysisof a fibrin clot.
 15. The method according to claim 13, wherein saidfibrinolytic agent is a plasminogen activator selected from the groupconsisting of anistreplase, urokinase, pro-urokinase, streptokinase,tissue plasminogen activator and mutants, or variants thereof, whichretain plasminogen activator activity.
 16. The method according to claim15, wherein said variants are selected from the group consisting ofvariants which have been chemically modified, variants which one or moreamino acids have been added, deleted or substituted, and variants withone or more modified functional domains.
 17. The method according toclaim 16, wherein said modified functional domains are added, deleted oraltered by combining the active site of one plasminogen activator orfibrin binding domain of another plasminogen activator or fibrin bindingmolecule.
 18. The method according to claim 6, wherein saidadministering is systemic administration of said compound to a subject.19. The method according to claim 18, wherein said systemicadministration is an administration selected from the group consistingof: injecting an injectable form of said compound; administering bymouth an oral form of said compound; applying to the skin a transdermalpatch or a transdermal pad containing said compound; administering aliquid/liquid suspension of said compound via nose drops or nasal spray;administering a nebulized liquid of said compound to oral ornasopharyngeal airways; administering rectally a suppository form ofsaid compound; administering vaginally said compound in dosage unitformulations containing conventional non-toxic pharmaceuticallyacceptable carriers, adjuvants and vehicles; administering said compoundintravitreally; and administering via intra-operative instillation agel, cream, powder, foam, crystals, liposomes, spray or liquidsuspension form of said compound; such that a therapeutically effectiveamount of said compound contacts the target platelets of said patientvia systemic absorption and circulation.
 20. The method according toclaim 18, wherein said systemic administration comprises infusion ofsaid compound to target platelets via a device selected from the groupconsisting of a pump catheter system and a continuous or selectiverelease device.
 21. The compound according to claim 1, wherein R₉ isaralkyl, in which the alkyl portion is a 2-carbon chain withunsaturation, and the aryl portion is a phenyl ring.
 22. Apharmaceutical formulation comprising the compound according to claim 21and a pharmaceutically acceptable carrier.
 23. A compound of FormulaIb″:

wherein: V═O; A=M; M=H or a pharmaceutically-acceptable inorganic ororganic counterion; D₁=O; Y′═H, OH, or OR₁; Z′=H, OH, or OR₂; with theproviso that at least one of Y′ and Z′ is OR₁ or OR₂; R₁ and R₂ areresidues which are linked directly to the 2′ and/or 3′ hydroxyls of thefuranose via a carbon atom according to Formula II,

wherein: O is the corresponding 2′- and/or 3′-oxygen of the respectivefuranose or carbocycle; C is a carbon atom; R₅, R⁶, and R₇ are H, alkyl,cycloalkyl, aralkyl, aryl, substituted aralkyl, or substituted aryl,such that the moiety defined according to Formula II is an ether; or R₅and R₆ are H, an alkyl, cycloalkyl, aralkyl, aryl, substituted aralkyl,or substituted aryl, and R₇ is alkoxy, cycloalkoxy, aralkyloxy, aryloxy,substituted aralkyloxy, or substituted aryloxy such that the moietydefined according to Formula II is an acyclic acetal or ketal; or R₅ andR₆ are taken together as oxygen or sulfur doubly bonded to C, and R₇ isalkyl, cycloalkyl, aralkyl, aryl, substituted aralkyl, or substitutedaryl, such that the moiety defined according to Formula II is an esteror thioester; or R₅ and R₆ are taken together as oxygen or sulfur doublybonded to C, and R₇ is amino or mono- or disubstituted amino, where thesubstituents are alkyl, cycloalkyl, aralkyl, aryl, substituted aralkyl,or substituted aryl, such that the moiety according to Formula II is acarbamate or thiocarbamate; or R₅ and R₆ are taken together as oxygen orsulfur doubly bonded to C, and R₇ is alkoxy, cycloalkoxy, aralkyloxy,aryloxy, substituted aralkyloxy, or substituted aryloxy, such that themoiety according to Formula II is a carbonate or thiocarbonate; or R₇ isnot present and R₅ and R₆ are taken together as oxygen or sulfur doublybonded to C and both the 2′- and 3′-oxygens of the respective furanoseor carbocycle are directly bound to C to form a cyclical carbonate orthiocarbonate; B′ is a purine residue according to general Formula IV:

wherein J=carbon; R₁₁ is absent; R₁₂ is hydrogen; R₁₃ is hydrogen; R₁₀is acylamino, according to Formula VI;

wherein: NH is the amino residue at the C-6 position in a purine; C is acarbon atom; W₁ is oxygen; and R₁₇ is amino or mono- or disubstitutedamino such that the moiety according to Formula VI is a urea.